Team:UT-Tokyo/LabNote

From 2011.igem.org

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=Parts List=
=Parts List=
 +
 +
Each part is represented by s number (e.g. #20 for lac promotor) in this notebook.
 +
By pointing the mouse on the number within notebook entires, you can find out the details of the part.
 +
 +
{| id="partslist"
{| id="partslist"
-
|+ Table 1. List of iGEM parts
+
|+ Table 1. List of iGEM parts used in our project
|-  
|-  
-
! Number !! Part !! Content !! Plasmid !! Length (bp)
+
! Number !! Reg.ID !! Content !! Plasmid !! Length (bp)
|-  
|-  
-
| #1 || J23119 || c.Promoter (Strong) || pSB1A2 || class="number" | 35
+
| #1 || [http://partsregistry.org/wiki/index.php?title=Part:BBa_J23119 J23119] || c.Promoter (Strong) || pSB1A2 || class="number" | 35
|-  
|-  
-
| #2 || J23118 || c.Promoter (Medium) || BBa_J61002 || class="number" |35
+
| #2 || [http://partsregistry.org/wiki/index.php?title=Part:BBa_J23118 J23118] || c.Promoter (Medium) || [http://partsregistry.org/wiki/index.php?title=Part:BBa_J61002 BBa_J61002] || class="number" |35
|-  
|-  
-
| #3 || B0032 || RBS || pSB1A2 || class="number" |13
+
| #3 || [http://partsregistry.org/wiki/index.php?title=Part:BBa_B0032 B0032] || RBS || pSB1A2 || class="number" |13
|-  
|-  
-
| #5 || B0014 || d.Ter || pSB1AK3 || class="number" |95
+
| #5 || [http://partsregistry.org/wiki/index.php?title=Part:BBa_B0014 B0014] || d.Ter || pSB1AK3 || class="number" |95
|-  
|-  
-
| #9 || E0240 || RBS-GFP-d.Ter || pSB1A2 || class="number" |876
+
| #9 || [http://partsregistry.org/wiki/index.php?title=Part:BBa_E0240 E0240] || RBS-GFP-d.Ter || pSB1A2 || class="number" |876
|-  
|-  
-
| #10 || E0040 || GFP || pSB1A2 || class="number" |681
+
| #10 || [http://partsregistry.org/wiki/index.php?title=Part:BBa_E0040 E0040] || GFP || pSB1A2 || class="number" |681
|-  
|-  
-
| #11 || E1010 || RFP || pSB2K3 || class="number" |723
+
| #11 || [http://partsregistry.org/wiki/index.php?title=Part:BBa_E1010 E1010] || RFP || pSB2K3 || class="number" |723
|-  
|-  
-
| #14 || I712019 || fLuc || pSB1AK8 || class="number" |1653
+
| #14 || [http://partsregistry.org/wiki/index.php?title=Part:BBa_I712019 I712019] || fLuc || pSB1AK8 || class="number" |1653
|-  
|-  
-
| #17 || J52008 || rLuc || pSB1AK3 || class="number" |936
+
| #17 || [http://partsregistry.org/wiki/index.php?title=Part:BBa_J52008 J52008] || rLuc || pSB1AK3 || class="number" |936
|-  
|-  
-
| #20 || R0011 || lacP || pSB1A2 || class="number" |55
+
| #20 || [http://partsregistry.org/wiki/index.php?title=Part:BBa_R0011 R0011] || lacP || pSB1A2 || class="number" |55
|-  
|-  
-
| #21 || C0012 || LacI || pSB1A2 || class="number" |1128
+
| #21 || [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0012 C0012] || LacI || pSB1A2 || class="number" |1128
|-  
|-  
-
| #22 || I712074 || pT7 || pSB1AK8 || class="number" |46
+
| #22 || [http://partsregistry.org/wiki/index.php?title=Part:BBa_I712074 I712074] || pT7 || pSB1AK8 || class="number" |46
|-  
|-  
-
| #23 || K145001 || T7 RNA Pol. || pSB1A2 || class="number" |2655
+
| #23 || [http://partsregistry.org/wiki/index.php?title=Part:BBa_K145001 K145001] || T7 RNA Pol. || pSB1A2 || class="number" |2655
|-  
|-  
-
| #24 || J22106 || recAp || pSB1A2 || class="number" |192
+
| #24 || [http://partsregistry.org/wiki/index.php?title=Part:BBa_J22106 J22106] || recAp || pSB1A2 || class="number" |192
|-  
|-  
-
| #27 || C0083 || AspA || pSB2K3 || class="number" |1518
+
| #27 || [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0083 C0083] || AspA || pSB2K3 || class="number" |1518
|-  
|-  
-
| #28 || K112808 || T4 phage lysis device (no promoter) || pSB1A2 || class="number" | 1785
+
| #28 || [http://partsregistry.org/wiki/index.php?title=Part:BBa_K112808 K112808] || T4 phage lysis device (no promoter) || pSB1A2 || class="number" | 1785
|-  
|-  
| #29 || - || CheZ || pSB1AK3 || class="number" | 728
| #29 || - || CheZ || pSB1AK3 || class="number" | 728
|-  
|-  
-
| #30 || K117000 || Lysis gene || pSB1A2 || class="number" | 144
+
| #30 || [http://partsregistry.org/wiki/index.php?title=Part:BBa_K117000 K117000] || Lysis gene || pSB1A2 || class="number" | 144
|-  
|-  
| #31 || - || LexA || pSB1AK3 || class="number" | 750
| #31 || - || LexA || pSB1AK3 || class="number" | 750
Line 51: Line 56:
| #35 || - || recNp ||  ||  
| #35 || - || recNp ||  ||  
|-  
|-  
-
| #36 || R0051 || cI-repressed promoter || pSB1A2 || class="number" | 49
+
| #36 || [http://partsregistry.org/wiki/index.php?title=Part:BBa_R0051 R0051] || cI-repressed promoter || pSB1A2 || class="number" | 49
|-  
|-  
-
| #37 || C0051 || cI repressor (LVA tagged) || pSB1A2 || \
+
| #37 || [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0051 C0051] || cI repressor (LVA tagged) || pSB1A2 || \
|-  
|-  
| #38 || - || RecA ||  ||  
| #38 || - || RecA ||  ||  
Line 149: Line 154:
-->
-->
-
=Lab Diary=
+
=Lab Notebook=
-
For convenience sake, each part (genes, promotors, etc) is represented by consecutive numbers (e.g. #20 for lac promotor).
+
-
By pointing the mouse on the part number in the construct, you can find out the details of the part.
+
<html>
<html>
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*Planting Negative control
*Planting Negative control
*Making frozen stock
*Making frozen stock
-
*Transformation(BBa_E0240, <nowiki>#</nowiki>3, B0015, <nowiki>#</nowiki>5, <nowiki>#</nowiki>10, ,J31000, ,J44000)
+
*Transformation(BBa_E0240, <nowiki>#</nowiki><html><span class="part_num">3</span></html>, B0015, <nowiki>#</nowiki><html><span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">10</span></html>, ,J31000, ,J44000)
<html></div></html>
<html></div></html>
Line 253: Line 256:
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/06/17(Fri)===
==='11/06/17(Fri)===
-
*Picking up colony(<nowiki>#</nowiki>3, B0015, B0040,<nowiki>#</nowiki>9, J31000, J44000)
+
*Picking up colony(<nowiki>#</nowiki><html><span class="part_num">3</span></html>, B0015, B0040,<nowiki>#</nowiki><html><span class="part_num">9</span></html>, J31000, J44000)
-
*Miniprep(<nowiki>#</nowiki>5)
+
*Miniprep(<nowiki>#</nowiki><html><span class="part_num">5</span></html>)
-
*Dissolution(<nowiki>#</nowiki>1, <nowiki>#</nowiki>2, K16500, <nowiki>#</nowiki>24)
+
*Dissolution(<nowiki>#</nowiki><html><span class="part_num">1</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span></html>, K16500, <nowiki>#</nowiki><html><span class="part_num">24</span></html>)
<html></div></html>
<html></div></html>
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/06/21(Tue)===
==='11/06/21(Tue)===
-
*Making frozen stock(<nowiki>#</nowiki>3, B0015, B0040, <nowiki>#</nowiki>9, J31000, J44000)
+
*Making frozen stock(<nowiki>#</nowiki><html><span class="part_num">3</span></html>, B0015, B0040, <nowiki>#</nowiki><html><span class="part_num">9</span></html>, J31000, J44000)
-
*Passafe culture(<nowiki>#</nowiki>9, J31000)
+
*Passafe culture(<nowiki>#</nowiki><html><span class="part_num">9</span></html>, J31000)
*Nanodrop again
*Nanodrop again
<html></div></html>
<html></div></html>
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<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/06/22(Wed)===
==='11/06/22(Wed)===
-
*Miniprep(<nowiki>#</nowiki>9, J31000)
+
*Miniprep(<nowiki>#</nowiki><html><span class="part_num">9</span></html>, J31000)
*Making competent cell
*Making competent cell
<html></div></html>
<html></div></html>
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*Digest(product of Miniprep 06/09 EPcut)
*Digest(product of Miniprep 06/09 EPcut)
*Electrophoresis
*Electrophoresis
-
*Defrost and transformation(BBa_E0030, E0020, <nowiki>#</nowiki>14, I712052,<nowiki>#</nowiki>17)
+
*Defrost and transformation(BBa_E0030, E0020, <nowiki>#</nowiki><html><span class="part_num">14</span></html>, I712052,<nowiki>#</nowiki><html><span class="part_num">17</span></html>)
*Making 1×TAE, agarose gel, LB medium, LB plate(amp),
*Making 1×TAE, agarose gel, LB medium, LB plate(amp),
<html></div></html>
<html></div></html>
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==='11/07/01(Fri)===
==='11/07/01(Fri)===
*Digest(EPcut)
*Digest(EPcut)
-
*Miniprep(BBa_E0030, E0020, <nowiki>#</nowiki>14, I712052, <nowiki>#</nowiki>17)
+
*Miniprep(BBa_E0030, E0020, <nowiki>#</nowiki><html><span class="part_num">14</span></html>, I712052, <nowiki>#</nowiki><html><span class="part_num">17</span></html>)
<html></div></html>
<html></div></html>
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==='11/07/05(Tue)===
==='11/07/05(Tue)===
*Electrophoresis(product of digest 07/01)
*Electrophoresis(product of digest 07/01)
-
*Digest(<nowiki>#</nowiki>9, <nowiki>#</nowiki>14, <nowiki>#</nowiki>17, <nowiki>#</nowiki>3, <nowiki>#</nowiki>5)
+
*Digest(<nowiki>#</nowiki><html><span class="part_num">9</span></html>, <nowiki>#</nowiki><html><span class="part_num">14</span></html>, <nowiki>#</nowiki><html><span class="part_num">17</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span></html>, <nowiki>#</nowiki><html><span class="part_num">5</span></html>)
*Making antibiotic stock(1000×)(Km,Cm)
*Making antibiotic stock(1000×)(Km,Cm)
<html></div></html>
<html></div></html>
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==='11/07/06(Wed)===
==='11/07/06(Wed)===
*Gel  extraction (pre)(BBa_I712052)
*Gel  extraction (pre)(BBa_I712052)
-
*Picking up colony(Ba_J23119, <nowiki>#</nowiki>2, <nowiki>#</nowiki>24, <nowiki>#</nowiki>10)
+
*Picking up colony(Ba_J23119, <nowiki>#</nowiki><html><span class="part_num">2</span></html>, <nowiki>#</nowiki><html><span class="part_num">24</span></html>, <nowiki>#</nowiki><html><span class="part_num">10</span></html>)
*Making agarose gel
*Making agarose gel
<html></div></html>
<html></div></html>
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<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/07/07(Thu)===
==='11/07/07(Thu)===
-
*Digest(BBa_E0240, <nowiki>#</nowiki>14, <nowiki>#</nowiki>17, <nowiki>#</nowiki>3, <nowiki>#</nowiki>5)
+
*Digest(BBa_E0240, <nowiki>#</nowiki><html><span class="part_num">14</span></html>, <nowiki>#</nowiki><html><span class="part_num">17</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span></html>, <nowiki>#</nowiki><html><span class="part_num">5</span></html>)
-
*Defrost(BBa_E1010, K325101, <nowiki>#</nowiki>20, <nowiki>#</nowiki>21, <nowiki>#</nowiki>22, <nowiki>#</nowiki>23)
+
*Defrost(BBa_E1010, K325101, <nowiki>#</nowiki><html><span class="part_num">20</span></html>, <nowiki>#</nowiki><html><span class="part_num">21</span></html>, <nowiki>#</nowiki><html><span class="part_num">22</span></html>, <nowiki>#</nowiki><html><span class="part_num">23</span></html>)
-
*Transformation(BBa_K145001, <nowiki>#</nowiki>20, <nowiki>#</nowiki>21, <nowiki>#</nowiki>22)
+
*Transformation(BBa_K145001, <nowiki>#</nowiki><html><span class="part_num">20</span></html>, <nowiki>#</nowiki><html><span class="part_num">21</span></html>, <nowiki>#</nowiki><html><span class="part_num">22</span></html>)
*Making LB plate(Cm,Km)
*Making LB plate(Cm,Km)
<html></div></html>
<html></div></html>
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<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/07/08(Fri)===
==='11/07/08(Fri)===
-
*Miniprep(BBa_J23119, <nowiki>#</nowiki>2,  <nowiki>#</nowiki>10, <nowiki>#</nowiki>24)
+
*Miniprep(BBa_J23119, <nowiki>#</nowiki><html><span class="part_num">2</span></html>,  <nowiki>#</nowiki><html><span class="part_num">10</span></html>, <nowiki>#</nowiki><html><span class="part_num">24</span></html>)
<html></div></html>
<html></div></html>
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/07/11(Mon)===
==='11/07/11(Mon)===
-
*Gel extraction(<nowiki>#</nowiki>9 <nowiki>#</nowiki>14, <nowiki>#</nowiki>17)
+
*Gel extraction(<nowiki>#</nowiki><html><span class="part_num">9</span></html> <nowiki>#</nowiki><html><span class="part_num">14</span></html>, <nowiki>#</nowiki><html><span class="part_num">17</span></html>)
-
*Transformation(<nowiki>#</nowiki>1, <nowiki>#</nowiki>11, <nowiki>#</nowiki>20, <nowiki>#</nowiki>21, K325101, <nowiki>#</nowiki>22, <nowiki>#</nowiki>23)
+
*Transformation(<nowiki>#</nowiki><html><span class="part_num">1</span></html>, <nowiki>#</nowiki><html><span class="part_num">11</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span></html>, <nowiki>#</nowiki><html><span class="part_num">21</span></html>, K325101, <nowiki>#</nowiki><html><span class="part_num">22</span></html>, <nowiki>#</nowiki><html><span class="part_num">23</span></html>)
<html></div></html>
<html></div></html>
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<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/07/13(Wed)===
==='11/07/13(Wed)===
-
*Frozen stock(<nowiki>#</nowiki>1(18A), <nowiki>#</nowiki>20(6G), <nowiki>#</nowiki>21(2O), <nowiki>#</nowiki>14(6N), <nowiki>#</nowiki>23(2F))
+
*Frozen stock(<nowiki>#</nowiki><html><span class="part_num">1</span></html>(18A), <nowiki>#</nowiki><html><span class="part_num">20</span></html>(6G), <nowiki>#</nowiki><html><span class="part_num">21</span></html>(2O), <nowiki>#</nowiki><html><span class="part_num">14</span></html>(6N), <nowiki>#</nowiki><html><span class="part_num">23</span></html>(2F))
-
*Picking up colony(<nowiki>#</nowiki>11)
+
*Picking up colony(<nowiki>#</nowiki><html><span class="part_num">11</span></html>)
-
*Gel extraction(B<nowiki>#</nowiki>3, <nowiki>#</nowiki>5)
+
*Gel extraction(B<nowiki>#</nowiki><html><span class="part_num">3</span></html>, <nowiki>#</nowiki><html><span class="part_num">5</span></html>)
-
*Digest (<nowiki>#</nowiki>2 SPcut)
+
*Digest (<nowiki>#</nowiki><html><span class="part_num">2</span></html> SPcut)
<html></div></html>
<html></div></html>
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/07/14(Thu)===
==='11/07/14(Thu)===
-
*Miniprep(<nowiki>#</nowiki>1, <nowiki>#</nowiki>11,  <nowiki>#</nowiki>20, <nowiki>#</nowiki>21, <nowiki>#</nowiki>22, <nowiki>#</nowiki>23)
+
*Miniprep(<nowiki>#</nowiki><html><span class="part_num">1</span></html>, <nowiki>#</nowiki><html><span class="part_num">11</span></html>,  <nowiki>#</nowiki><html><span class="part_num">20</span></html>, <nowiki>#</nowiki><html><span class="part_num">21</span></html>, <nowiki>#</nowiki><html><span class="part_num">22</span></html>, <nowiki>#</nowiki><html><span class="part_num">23</span></html>)
-
*Electrophoresis(<nowiki>#</nowiki>2)
+
*Electrophoresis(<nowiki>#</nowiki><html><span class="part_num">2</span></html>)
-
*Gel extraction(<nowiki>#</nowiki>2, <nowiki>#</nowiki>3, <nowiki>#</nowiki>5)
+
*Gel extraction(<nowiki>#</nowiki><html><span class="part_num">2</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span></html>, <nowiki>#</nowiki><html><span class="part_num">5</span></html>)
-
*Passafe(<nowiki>#</nowiki>11)
+
*Passafe(<nowiki>#</nowiki><html><span class="part_num">11</span></html>)
*Making LB+amp
*Making LB+amp
<html></div></html>
<html></div></html>
Line 358: Line 361:
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/07/15(Fri)===
==='11/07/15(Fri)===
-
*Ligation(<nowiki>#</nowiki>2-<nowiki>#</nowiki>9, <nowiki>#</nowiki>2-<nowiki>#</nowiki>3, <nowiki>#</nowiki>14-<nowiki>#</nowiki>5, <nowiki>#</nowiki>17-<nowiki>#</nowiki>5)
+
*Ligation(<nowiki>#</nowiki><html><span class="part_num">2</span>-</html><nowiki>#</nowiki><html><span class="part_num">9</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span></html>, <nowiki>#</nowiki><html><span class="part_num">14</span>-</html><nowiki>#</nowiki><html><span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">17</span>-</html><nowiki>#</nowiki><html><span class="part_num">5</span></html>)
-
*Transformation(<nowiki>#</nowiki>2-<nowiki>#</nowiki>9, <nowiki>#</nowiki>2-<nowiki>#</nowiki>3, <nowiki>#</nowiki>14-<nowiki>#</nowiki>5, <nowiki>#</nowiki>17-<nowiki>#</nowiki>5)
+
*Transformation(<nowiki>#</nowiki><html><span class="part_num">2</span>-</html><nowiki>#</nowiki><html><span class="part_num">9</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span></html>, <nowiki>#</nowiki><html><span class="part_num">14</span>-</html><nowiki>#</nowiki><html><span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">17</span>-</html><nowiki>#</nowiki><html><span class="part_num">5</span></html>)
*Dispensing Ligation Buffer
*Dispensing Ligation Buffer
-
*Frozen stock(<nowiki>#</nowiki>11)
+
*Frozen stock(<nowiki>#</nowiki><html><span class="part_num">11</span></html>)
<html></div></html>
<html></div></html>
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*Making Master plate
*Making Master plate
*Testing product of Miniprep
*Testing product of Miniprep
-
*Transformation (<nowiki>#</nowiki>2, <nowiki>#</nowiki>27, <nowiki>#</nowiki>28)
+
*Transformation (<nowiki>#</nowiki><html><span class="part_num">2</span></html>, <nowiki>#</nowiki><html><span class="part_num">27</span></html>, <nowiki>#</nowiki><html><span class="part_num">28</span></html>)
<html></div></html>
<html></div></html>
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/07/21(Thu)===
==='11/07/21(Thu)===
-
*Miniprep(<nowiki>#</nowiki>2-3, <nowiki>#</nowiki>2-9, <nowiki>#</nowiki>14-5, <nowiki>#</nowiki>17-5, <nowiki>#</nowiki>11, <nowiki>#</nowiki>20)
+
*Miniprep(<nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">9</span></html>, <nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">11</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span></html>)
*Testing product
*Testing product
*Digest
*Digest
-
**SPcut : <nowiki>#</nowiki>20, <nowiki>#</nowiki>2-3
+
**SPcut : <nowiki>#</nowiki><html><span class="part_num">20</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span></html>
-
**XPcut : <nowiki>#</nowiki>10, <nowiki>#</nowiki>11, <nowiki>#</nowiki>14-5, <nowiki>#</nowiki>17-5
+
**XPcut : <nowiki>#</nowiki><html><span class="part_num">10</span></html>, <nowiki>#</nowiki><html><span class="part_num">11</span></html>, <nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">17</span>-<span class="part_num">5</span></html>
*Making TB
*Making TB
<html></div></html>
<html></div></html>
Line 391: Line 394:
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/07/22(Fri)===
==='11/07/22(Fri)===
-
*Frozen stock(<nowiki>#</nowiki>2, <nowiki>#</nowiki>2-3, <nowiki>#</nowiki>2-9, <nowiki>#</nowiki>14-5, <nowiki>#</nowiki>17-5)
+
*Frozen stock(<nowiki>#</nowiki><html><span class="part_num">2</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">9</span></html>, <nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">17</span>-<span class="part_num">5</span></html>)
-
*Electrophoresis(<nowiki>#</nowiki>20_SP, <nowiki>#</nowiki>2-3_SP, <nowiki>#</nowiki>10_XP, <nowiki>#</nowiki>11_XP, <nowiki>#</nowiki>14-5_XP, <nowiki>#</nowiki>17-5_XP)
+
*Electrophoresis(<nowiki>#</nowiki><html><span class="part_num">20</span></html>_SP, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span></html>_SP, <nowiki>#</nowiki><html><span class="part_num">10</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">11</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">17</span>-<span class="part_num">5</span></html>_XP)
*Digest
*Digest
-
**SPcut : <nowiki>#</nowiki>20
+
**SPcut : <nowiki>#</nowiki><html><span class="part_num">20</span></html>
-
**XPcut : <nowiki>#</nowiki>10, <nowiki>#</nowiki>11, <nowiki>#</nowiki>14-5, <nowiki>#</nowiki>17-5
+
**XPcut : <nowiki>#</nowiki><html><span class="part_num">10</span></html>, <nowiki>#</nowiki><html><span class="part_num">11</span></html>, <nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">17</span>-<span class="part_num">5</span></html>
-
*Transformation(<nowiki>#</nowiki>27, <nowiki>#</nowiki>28, product of Miniprep 07/20)
+
*Transformation(<nowiki>#</nowiki><html><span class="part_num">27</span></html>, <nowiki>#</nowiki><html><span class="part_num">28</span></html>, product of Miniprep 07/20)
<html></div></html>
<html></div></html>
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/07/25(Mon)===
==='11/07/25(Mon)===
-
*Colony check(<nowiki>#</nowiki>2, <nowiki>#</nowiki>2-9)
+
*Colony check(<nowiki>#</nowiki><html><span class="part_num">2</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">9</span></html>)
-
*Gel Extraction(<nowiki>#</nowiki>20_SP, <nowiki>#</nowiki>10_XP, <nowiki>#</nowiki>11_XP, <nowiki>#</nowiki>14-5_XP, <nowiki>#</nowiki>17-5_XP)
+
*Gel Extraction(<nowiki>#</nowiki><html><span class="part_num">20</span></html>_SP, <nowiki>#</nowiki><html><span class="part_num">10</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">11</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">17</span>-<span class="part_num">5</span></html>_XP)
-
*Ligation and Transformation(<nowiki>#</nowiki>20-<nowiki>#</nowiki>9, <nowiki>#</nowiki>3-<nowiki>#</nowiki>11, <nowiki>#</nowiki>3-<nowiki>#</nowiki>14-5, <nowiki>#</nowiki>3-<nowiki>#</nowiki>17-5)
+
*Ligation and Transformation(<nowiki>#</nowiki><html><span class="part_num">20</span>-</html><nowiki>#</nowiki><html><span class="part_num">9</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">11</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">17</span>-<span class="part_num">5</span></html>)
*Defrost Primer(200×, 10×)
*Defrost Primer(200×, 10×)
*Dispensing PCR Mix
*Dispensing PCR Mix
Line 410: Line 413:
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/07/26(Tue)===
==='11/07/26(Tue)===
-
*Picking up colony and making master plate(<nowiki>#</nowiki>20-9, <nowiki>#</nowiki>3-11, <nowiki>#</nowiki>3-14-5, <nowiki>#</nowiki>3-17-5)
+
*Picking up colony and making master plate(<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">9</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
-
*Colony PCR(<nowiki>#</nowiki>20-9, <nowiki>#</nowiki>3-11, <nowiki>#</nowiki>3-14-5, <nowiki>#</nowiki>3-17-5)
+
*Colony PCR(<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">9</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
-
*Digest(<nowiki>#</nowiki>14-5 XPcut,Ecut, <nowiki>#</nowiki>22 SPcut, <nowiki>#</nowiki>23 XPcut)
+
*Digest(<nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html> XPcut,Ecut, <nowiki>#</nowiki><html><span class="part_num">22</span></html> SPcut, <nowiki>#</nowiki><html><span class="part_num">23</span></html> XPcut)
-
*Ligation(<nowiki>#</nowiki>3-<nowiki>#</nowiki>14-5(Re), <nowiki>#</nowiki>3(Negative control))
+
*Ligation(<nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>(Re), <nowiki>#</nowiki><html><span class="part_num">3</span></html>(Negative control))
-
*Transformation(<nowiki>#</nowiki>15, <nowiki>#</nowiki>27, <nowiki>#</nowiki>28, <nowiki>#</nowiki>30, <nowiki>#</nowiki>3-14-5(Re), <nowiki>#</nowiki>3(Negative control))
+
*Transformation(<nowiki>#</nowiki><html><span class="part_num">15</span></html>, <nowiki>#</nowiki><html><span class="part_num">27</span></html>, <nowiki>#</nowiki><html><span class="part_num">28</span></html>, <nowiki>#</nowiki><html><span class="part_num">30</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>(Re), <nowiki>#</nowiki><html><span class="part_num">3</span></html>(Negative control))
*Making reagent for TB(KOH, CaCl2, KCl, MnCl2)
*Making reagent for TB(KOH, CaCl2, KCl, MnCl2)
<html></div></html>
<html></div></html>
Line 420: Line 423:
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/07/27(Wed)===
==='11/07/27(Wed)===
-
*Miniprep(<nowiki>#</nowiki>1, <nowiki>#</nowiki>2, <nowiki>#</nowiki>3, <nowiki>#</nowiki>10, <nowiki>#</nowiki>22, <nowiki>#</nowiki>24, <nowiki>#</nowiki>20-9, <nowiki>#</nowiki>3-11, <nowiki>#</nowiki>3-17-5)
+
*Miniprep(<nowiki>#</nowiki><html><span class="part_num">1</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span></html>, <nowiki>#</nowiki><html><span class="part_num">10</span></html>, <nowiki>#</nowiki><html><span class="part_num">22</span></html>, <nowiki>#</nowiki><html><span class="part_num">24</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">9</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
*Electrophoresis
*Electrophoresis
<html></div></html>
<html></div></html>
Line 426: Line 429:
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/07/28(Thu)===
==='11/07/28(Thu)===
-
*Gel extraction(<nowiki>#</nowiki>14-5_XP, <nowiki>#</nowiki>22_SP, <nowiki>#</nowiki>23_XP)
+
*Gel extraction(<nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">22</span></html>_SP, <nowiki>#</nowiki><html><span class="part_num">23</span></html>_XP)
*Digest
*Digest
-
**SPcut : <nowiki>#</nowiki>1,<nowiki>#</nowiki>2, <nowiki>#</nowiki>3, <nowiki>#</nowiki>24
+
**SPcut : <nowiki>#</nowiki><html><span class="part_num">1</span></html>,<nowiki>#</nowiki><html><span class="part_num">2</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span></html>, <nowiki>#</nowiki><html><span class="part_num">24</span></html>
-
**EScut : <nowiki>#</nowiki>3-11
+
**EScut : <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span></html>
-
**XPcut : <nowiki>#</nowiki>3-17-5
+
**XPcut : <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>
-
*Ligation(<nowiki>#</nowiki>3-<nowiki>#</nowiki>14-5)
+
*Ligation(<nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>)
*Making TB bugger, LB plate, 0.3% LB plate
*Making TB bugger, LB plate, 0.3% LB plate
<html></div></html>
<html></div></html>
Line 438: Line 441:
==='11/07/29(Fri)===
==='11/07/29(Fri)===
*Cloning(lexA, cheZ)
*Cloning(lexA, cheZ)
-
*Colony PCR(<nowiki>#</nowiki>3-<nowiki>#</nowiki>14-5)
+
*Colony PCR(<nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>)
-
*Gel extraction(<nowiki>#</nowiki>1_SP, <nowiki>#</nowiki>2_SP, <nowiki>#</nowiki>3_SP, <nowiki>#</nowiki>24_SP, <nowiki>#</nowiki>3-11_ES, <nowiki>#</nowiki>3-17-5_XP, lexA, cheZ PCR product)
+
*Gel extraction(<nowiki>#</nowiki><html><span class="part_num">1</span></html>_SP, <nowiki>#</nowiki><html><span class="part_num">2</span></html>_SP, <nowiki>#</nowiki><html><span class="part_num">3</span></html>_SP, <nowiki>#</nowiki><html><span class="part_num">24</span></html>_SP, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span></html>_ES, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_XP, lexA, cheZ PCR product)
-
*Miniprep(<nowiki>#</nowiki>30)
+
*Miniprep(<nowiki>#</nowiki><html><span class="part_num">30</span></html>)
*Digest(lexA, cheZ PCR product XP cut)
*Digest(lexA, cheZ PCR product XP cut)
<html></div></html>
<html></div></html>
Line 449: Line 452:
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/08/01(Mon)===
==='11/08/01(Mon)===
-
*Nanodrop(<nowiki>#</nowiki>3-11(1), <nowiki>#</nowiki>1, <nowiki>#</nowiki>3, <nowiki>#</nowiki>24, <nowiki>#</nowiki>2, <nowiki>#</nowiki>3-17-5(4)M<nowiki>#</nowiki>30)
+
*Nanodrop(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span></html>(1), <nowiki>#</nowiki><html><span class="part_num">1</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span></html>, <nowiki>#</nowiki><html><span class="part_num">24</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>(4)M<nowiki>#</nowiki><html><span class="part_num">30</span></html>)
-
*Digest(<nowiki>#</nowiki>3 SPcut, <nowiki>#</nowiki>5 EXcut, <nowiki>#</nowiki>9 XPcut, <nowiki>#</nowiki>30 XP cut)
+
*Digest(<nowiki>#</nowiki><html><span class="part_num">3</span></html> SPcut, <nowiki>#</nowiki><html><span class="part_num">5</span></html> EXcut, <nowiki>#</nowiki><html><span class="part_num">9</span></html> XPcut, <nowiki>#</nowiki><html><span class="part_num">30</span></html> XP cut)
*Cloning(lexA, cheZ)
*Cloning(lexA, cheZ)
-
*Gel extraction(<nowiki>#</nowiki>3, <nowiki>#</nowiki>9, <nowiki>#</nowiki>30, pSB1A2)
+
*Gel extraction(<nowiki>#</nowiki><html><span class="part_num">3</span></html>, <nowiki>#</nowiki><html><span class="part_num">9</span></html>, <nowiki>#</nowiki><html><span class="part_num">30</span></html>, pSB1A2)
-
*Ligation(<nowiki>#</nowiki>2-<nowiki>#</nowiki>3-17-5, <nowiki>#</nowiki>3-11-<nowiki>#</nowiki>5, <nowiki>#</nowiki>14-<nowiki>#</nowiki>5, <nowiki>#</nowiki>3-<nowiki>#</nowiki>23, <nowiki>#</nowiki>22-<nowiki>#</nowiki>9, <nowiki>#</nowiki>3-<nowiki>#</nowiki>30)
+
*Ligation(<nowiki>#</nowiki><html><span class="part_num">2</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-</html><nowiki>#</nowiki><html><span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">14</span>-</html><nowiki>#</nowiki><html><span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">23</span></html>, <nowiki>#</nowiki><html><span class="part_num">22</span>-</html><nowiki>#</nowiki><html><span class="part_num">9</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">30</span></html>)
-
*Transformation(<nowiki>#</nowiki>2-<nowiki>#</nowiki>3-17-5, <nowiki>#</nowiki>3-11-<nowiki>#</nowiki>5, <nowiki>#</nowiki>14-<nowiki>#</nowiki>5, <nowiki>#</nowiki>3-<nowiki>#</nowiki>23, <nowiki>#</nowiki>22-<nowiki>#</nowiki>9, <nowiki>#</nowiki>3-<nowiki>#</nowiki>30, <nowiki>#</nowiki>27(IPTG), <nowiki>#</nowiki>28)
+
*Transformation(<nowiki>#</nowiki><html><span class="part_num">2</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-</html><nowiki>#</nowiki><html><span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">14</span>-</html><nowiki>#</nowiki><html><span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">23</span></html>, <nowiki>#</nowiki><html><span class="part_num">22</span>-</html><nowiki>#</nowiki><html><span class="part_num">9</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">30</span></html>, <nowiki>#</nowiki><html><span class="part_num">27</span></html>(IPTG), <nowiki>#</nowiki><html><span class="part_num">28</span></html>)
<html></div></html>
<html></div></html>
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/08/02(Tue)===
==='11/08/02(Tue)===
-
*Colony PCR(<nowiki>#</nowiki>2-<nowiki>#</nowiki>3-17-5, <nowiki>#</nowiki>3-11-<nowiki>#</nowiki>5, <nowiki>#</nowiki>14-<nowiki>#</nowiki>5, <nowiki>#</nowiki>3-<nowiki>#</nowiki>23, <nowiki>#</nowiki>22-<nowiki>#</nowiki>9)
+
*Colony PCR(<nowiki>#</nowiki><html><span class="part_num">2</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-</html><nowiki>#</nowiki><html><span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">14</span>-</html><nowiki>#</nowiki><html><span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">23</span></html>, <nowiki>#</nowiki><html><span class="part_num">22</span>-</html><nowiki>#</nowiki><html><span class="part_num">9</span></html>)
-
*Cloning(PCR)(<nowiki>#</nowiki>27, <nowiki>#</nowiki>28)
+
*Cloning(PCR)(<nowiki>#</nowiki><html><span class="part_num">27</span></html>, <nowiki>#</nowiki><html><span class="part_num">28</span></html>)
*Gel extraction
*Gel extraction
-
*PCR_2nd(lexA, cheZ, <nowiki>#</nowiki>27, <nowiki>#</nowiki>28)
+
*PCR_2nd(lexA, cheZ, <nowiki>#</nowiki><html><span class="part_num">27</span></html>, <nowiki>#</nowiki><html><span class="part_num">28</span></html>)
-
*Digest (<nowiki>#</nowiki>30 again)
+
*Digest (<nowiki>#</nowiki><html><span class="part_num">30</span></html> again)
*Making competent cell(cheZ-, Wild Type)
*Making competent cell(cheZ-, Wild Type)
*Miniprep
*Miniprep
Line 470: Line 473:
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/08/04(Thu)===
==='11/08/04(Thu)===
-
*Electrophoresis(product of ligation <nowiki>#</nowiki>2-3-17-5(2), <nowiki>#</nowiki>3-11-5(1,2), <nowiki>#</nowiki>14-5(2), <nowiki>#</nowiki>22-9(1,2,3))
+
*Electrophoresis(product of ligation <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>(2), <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>(1,2), <nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>(2), <nowiki>#</nowiki><html><span class="part_num">22</span>-<span class="part_num">9</span></html>(1,2,3))
-
*Frozen stock(<nowiki>#</nowiki>2-3-17-5(2), <nowiki>#</nowiki>3-11-5(1,2), <nowiki>#</nowiki>14-5(2), <nowiki>#</nowiki>22-9(1,2))
+
*Frozen stock(<nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>(2), <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>(1,2), <nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>(2), <nowiki>#</nowiki><html><span class="part_num">22</span>-<span class="part_num">9</span></html>(1,2))
*Making Gel
*Making Gel
-
*Digest(<nowiki>#</nowiki>23 XPcut)
+
*Digest(<nowiki>#</nowiki><html><span class="part_num">23</span></html> XPcut)
-
*IPTG induct(<nowiki>#</nowiki>20-9)
+
*IPTG induct(<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">9</span></html>)
<html></div></html>
<html></div></html>
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/08/05(Fri)===
==='11/08/05(Fri)===
-
*Digest(<nowiki>#</nowiki>14-5(2) XPcut, <nowiki>#</nowiki>3-11-5(1) XPcut)
+
*Digest(<nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>(2) XPcut, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>(1) XPcut)
-
*Cloning(PCR)(<nowiki>#</nowiki>27, <nowiki>#</nowiki>28, lexA, cheZ)
+
*Cloning(PCR)(<nowiki>#</nowiki><html><span class="part_num">27</span></html>, <nowiki>#</nowiki><html><span class="part_num">28</span></html>, lexA, cheZ)
<html></div></html>
<html></div></html>
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/08/08(Mon)===
==='11/08/08(Mon)===
-
*Defrost Frozen stock, Miniprep, Nanodrop and Electrophoresis(<nowiki>#</nowiki>5, <nowiki>#</nowiki>20, <nowiki>#</nowiki>23, <nowiki>#</nowiki>30)
+
*Defrost Frozen stock, Miniprep, Nanodrop and Electrophoresis(<nowiki>#</nowiki><html><span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span></html>, <nowiki>#</nowiki><html><span class="part_num">23</span></html>, <nowiki>#</nowiki><html><span class="part_num">30</span></html>)
*Making Gel
*Making Gel
*Digest
*Digest
-
**XPcut : <nowiki>#</nowiki>27, <nowiki>#</nowiki>28, cheZ, lexA, <nowiki>#</nowiki>3-11-5(1), <nowiki>#</nowiki>23,<nowiki>#</nowiki>30
+
**XPcut : <nowiki>#</nowiki><html><span class="part_num">27</span></html>, <nowiki>#</nowiki><html><span class="part_num">28</span></html>, cheZ, lexA, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>(1), <nowiki>#</nowiki><html><span class="part_num">23</span></html>,<nowiki>#</nowiki><html><span class="part_num">30</span></html>
-
**EScut : <nowiki>#</nowiki>6
+
**EScut : <nowiki>#</nowiki><html><span class="part_num">6</span></html>
-
**EXcut : <nowiki>#</nowiki>7
+
**EXcut : <nowiki>#</nowiki><html><span class="part_num">7</span></html>
-
**SPcut : <nowiki>#</nowiki>7,<nowiki>#</nowiki>20
+
**SPcut : <nowiki>#</nowiki><html><span class="part_num">7</span></html>,<nowiki>#</nowiki><html><span class="part_num">20</span></html>
-
*Defrost Frozen stock(<nowiki>#</nowiki>5, <nowiki>#</nowiki>14-5(2), <nowiki>#</nowiki>3-11-5(1))
+
*Defrost Frozen stock(<nowiki>#</nowiki><html><span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>(2), <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>(1))
<html></div></html>
<html></div></html>
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/08/09(Tue)===
==='11/08/09(Tue)===
-
*Miniprep, Electrophoresis and Digest(<nowiki>#</nowiki>5, <nowiki>#</nowiki>14-5(2), <nowiki>#</nowiki>3-11-5(1))
+
*Miniprep, Electrophoresis and Digest(<nowiki>#</nowiki><html><span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>(2), <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>(1))
-
*Gel extraction and Nanodrop(<nowiki>#</nowiki>6, <nowiki>#</nowiki>27, <nowiki>#</nowiki>8, cheZ, lexA③, <nowiki>#</nowiki>3-11-5(1), <nowiki>#</nowiki>23, <nowiki>#</nowiki>7_EX, <nowiki>#</nowiki>20, <nowiki>#</nowiki>7_SP)
+
*Gel extraction and Nanodrop(<nowiki>#</nowiki><html><span class="part_num">6</span></html>, <nowiki>#</nowiki><html><span class="part_num">27</span></html>, <nowiki>#</nowiki><html><span class="part_num">8</span></html>, cheZ, lexA③, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>(1), <nowiki>#</nowiki><html><span class="part_num">23</span></html>, <nowiki>#</nowiki><html><span class="part_num">7</span></html>_EX, <nowiki>#</nowiki><html><span class="part_num">20</span></html>, <nowiki>#</nowiki><html><span class="part_num">7</span></html>_SP)
-
*Ligation(<nowiki>#</nowiki>27-psB1A2, <nowiki>#</nowiki>3-27, <nowiki>#</nowiki>20-28, <nowiki>#</nowiki>28-psB1A2, <nowiki>#</nowiki>3-29, <nowiki>#</nowiki>29-psB1A2, <nowiki>#</nowiki>31-psB1A2, <nowiki>#</nowiki>3-<nowiki>#</nowiki>23, <nowiki>#</nowiki>1-<nowiki>#</nowiki>3-11-5(1), <nowiki>#</nowiki>2-<nowiki>#</nowiki>3-11-5(1), <nowiki>#</nowiki>24-<nowiki>#</nowiki>3-11-5(1), <nowiki>#</nowiki>3-<nowiki>#</nowiki>30)
+
*Ligation(<nowiki>#</nowiki><html><span class="part_num">27</span>-</html>psB1A2, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">28</span></html>, <nowiki>#</nowiki><html><span class="part_num">28</span>-</html>psB1A2, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span></html>, <nowiki>#</nowiki><html><span class="part_num">29</span>-</html>psB1A2, <nowiki>#</nowiki><html><span class="part_num">31</span>-</html>psB1A2, <nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">23</span></html>, <nowiki>#</nowiki><html><span class="part_num">1</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>(1), <nowiki>#</nowiki><html><span class="part_num">2</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>(1), <nowiki>#</nowiki><html><span class="part_num">24</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>(1), <nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">30</span></html>)
*Making LB plate
*Making LB plate
<html></div></html>
<html></div></html>
Line 505: Line 508:
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/08/10(Wed)===
==='11/08/10(Wed)===
-
*Colony PCR(<nowiki>#</nowiki>27-pSB1A2, <nowiki>#</nowiki>3-27, <nowiki>#</nowiki>20-28, <nowiki>#</nowiki>28-psB1A2, <nowiki>#</nowiki>3-29, <nowiki>#</nowiki>29-psB1A2, <nowiki>#</nowiki>31-pSB1A2, <nowiki>#</nowiki>3-<nowiki>#</nowiki>23, <nowiki>#</nowiki>1-<nowiki>#</nowiki>3-11-5(1), <nowiki>#</nowiki>2-<nowiki>#</nowiki>3-11-5(1), <nowiki>#</nowiki>24-<nowiki>#</nowiki>3-11-5(1), <nowiki>#</nowiki>3-<nowiki>#</nowiki>30))
+
*Colony PCR(<nowiki>#</nowiki><html><span class="part_num">27</span>-</html>pSB1A2, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">28</span></html>, <nowiki>#</nowiki><html><span class="part_num">28</span>-</html>psB1A2, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span></html>, <nowiki>#</nowiki><html><span class="part_num">29</span>-</html>psB1A2, <nowiki>#</nowiki><html><span class="part_num">31</span>-</html>pSB1A2, <nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">23</span></html>, <nowiki>#</nowiki><html><span class="part_num">1</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>(1), <nowiki>#</nowiki><html><span class="part_num">2</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>(1), <nowiki>#</nowiki><html><span class="part_num">24</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>(1), <nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">30</span></html>))
*Cloning(lexA, cheZ)
*Cloning(lexA, cheZ)
*Digest
*Digest
-
**EScut : <nowiki>#</nowiki>6
+
**EScut : <nowiki>#</nowiki><html><span class="part_num">6</span></html>
-
**EXcut : <nowiki>#</nowiki>7
+
**EXcut : <nowiki>#</nowiki><html><span class="part_num">7</span></html>
-
**SPcut : <nowiki>#</nowiki>1, <nowiki>#</nowiki>2, <nowiki>#</nowiki>3, <nowiki>#</nowiki>7, <nowiki>#</nowiki>20
+
**SPcut : <nowiki>#</nowiki><html><span class="part_num">1</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span></html>, <nowiki>#</nowiki><html><span class="part_num">7</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span></html>
-
**XPcut : <nowiki>#</nowiki>23
+
**XPcut : <nowiki>#</nowiki><html><span class="part_num">23</span></html>
*Defrost Primer
*Defrost Primer
<html></div></html>
<html></div></html>
Line 517: Line 520:
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/08/11(Thu)===
==='11/08/11(Thu)===
-
*Gel extraction(<nowiki>#</nowiki>30, <nowiki>#</nowiki>14-5(insert,vector), <nowiki>#</nowiki>5, <nowiki>#</nowiki>3-11-5, cheZ, lexA)
+
*Gel extraction(<nowiki>#</nowiki><html><span class="part_num">30</span></html>, <nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>(insert,vector), <nowiki>#</nowiki><html><span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>, cheZ, lexA)
*Cloning(sulAp, uvrAp)
*Cloning(sulAp, uvrAp)
-
*Miniprep(<nowiki>#</nowiki>3, <nowiki>#</nowiki>20-28, <nowiki>#</nowiki>31, <nowiki>#</nowiki>24-3-11-5, <nowiki>#</nowiki>3-30)
+
*Miniprep(<nowiki>#</nowiki><html><span class="part_num">3</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">28</span></html>, <nowiki>#</nowiki><html><span class="part_num">31</span></html>, <nowiki>#</nowiki><html><span class="part_num">24</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">30</span></html>)
<html></div></html>
<html></div></html>
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/08/12(Fri)===
==='11/08/12(Fri)===
-
*Colony PCR(<nowiki>#</nowiki>3-14-5(2), <nowiki>#</nowiki>6-5, <nowiki>#</nowiki>3-23)
+
*Colony PCR(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>(2), <nowiki>#</nowiki><html><span class="part_num">6</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">23</span></html>)
*Cloning(nested-PCR)(uvrAp, sulAp)
*Cloning(nested-PCR)(uvrAp, sulAp)
*Check Electrophoresis
*Check Electrophoresis
-
**8/11 PCR product:<nowiki>#</nowiki>27, <nowiki>#</nowiki>28
+
**8/11 PCR product:<nowiki>#</nowiki><html><span class="part_num">27</span></html>, <nowiki>#</nowiki><html><span class="part_num">28</span></html>
-
**Miniprep product:<nowiki>#</nowiki>3-14-5(2), <nowiki>#</nowiki>5
+
**Miniprep product:<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>(2), <nowiki>#</nowiki><html><span class="part_num">5</span></html>
-
*Digest(<nowiki>#</nowiki>5 EXcut, <nowiki>#</nowiki>14 EScut)
+
*Digest(<nowiki>#</nowiki><html><span class="part_num">5</span></html> EXcut, <nowiki>#</nowiki><html><span class="part_num">14</span></html> EScut)
*Gel extraction
*Gel extraction
-
**8/11 digest product : <nowiki>#</nowiki>5, <nowiki>#</nowiki>14-5, <nowiki>#</nowiki>3-30, cheZ, lexA
+
**8/11 digest product : <nowiki>#</nowiki><html><span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">30</span></html>, cheZ, lexA
-
**8/11 PCR product : <nowiki>#</nowiki>27, <nowiki>#</nowiki>28
+
**8/11 PCR product : <nowiki>#</nowiki><html><span class="part_num">27</span></html>, <nowiki>#</nowiki><html><span class="part_num">28</span></html>
**8/12 nested-PCR product : sulAp, uvrAp
**8/12 nested-PCR product : sulAp, uvrAp
-
*Colony PCR (re) (<nowiki>#</nowiki>13-14-5)
+
*Colony PCR (re) (<nowiki>#</nowiki><html><span class="part_num">13</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>)
-
*Miniprep(ligation product : <nowiki>#</nowiki>6-5, <nowiki>#</nowiki>3-23)
+
*Miniprep(ligation product : <nowiki>#</nowiki><html><span class="part_num">6</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">23</span></html>)
*Cloning(hotstart-PCR: uvrBp , cheZ)
*Cloning(hotstart-PCR: uvrBp , cheZ)
*Digest
*Digest
-
**XPcut:<nowiki>#</nowiki>27, <nowiki>#</nowiki>28, <nowiki>#</nowiki>6-5
+
**XPcut:<nowiki>#</nowiki><html><span class="part_num">27</span></html>, <nowiki>#</nowiki><html><span class="part_num">28</span></html>, <nowiki>#</nowiki><html><span class="part_num">6</span>-<span class="part_num">5</span></html>
-
**EScut: sulAp, uvrAp, <nowiki>#</nowiki>3-23
+
**EScut: sulAp, uvrAp, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">23</span></html>
<html></div></html>
<html></div></html>
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/08/15(Mon)===
==='11/08/15(Mon)===
-
*Gel extraction(from previous digest products(8/12) : <nowiki>#</nowiki>27, <nowiki>#</nowiki>28, <nowiki>#</nowiki>33, <nowiki>#</nowiki>34, <nowiki>#</nowiki>4, <nowiki>#</nowiki>3-23)
+
*Gel extraction(from previous digest products(8/12) : <nowiki>#</nowiki><html><span class="part_num">27</span></html>, <nowiki>#</nowiki><html><span class="part_num">28</span></html>, <nowiki>#</nowiki><html><span class="part_num">33</span></html>, <nowiki>#</nowiki><html><span class="part_num">34</span></html>, <nowiki>#</nowiki><html><span class="part_num">4</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">23</span></html>)
*Digest
*Digest
**XPcut: cheZ, uvrBp(PCR products)  
**XPcut: cheZ, uvrBp(PCR products)  
-
**EScut: <nowiki>#</nowiki>3-11-5 (to obtain pSB1AK3 EScut)
+
**EScut: <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html> (to obtain pSB1AK3 EScut)
*Assay of Cell lysis construction(pLuc-Lysis device) inducted by IPTG
*Assay of Cell lysis construction(pLuc-Lysis device) inducted by IPTG
*Making Lysis buffer, Agarose gel
*Making Lysis buffer, Agarose gel
-
*Gel extraction(from digest products XPcut: cheZ, uvrBp(PCR products) EScut: <nowiki>#</nowiki>3-11-5 to obtain pSB1AK3 EScut)
+
*Gel extraction(from digest products XPcut: cheZ, uvrBp(PCR products) EScut: <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html> to obtain pSB1AK3 EScut)
-
*Ligation(<nowiki>#</nowiki>3-<nowiki>#</nowiki>14-5, <nowiki>#</nowiki>24-<nowiki>#</nowiki>3-17-5, <nowiki>#</nowiki>24-<nowiki>#</nowiki>3-11-5, <nowiki>#</nowiki>20-<nowiki>#</nowiki>28, <nowiki>#</nowiki>3-<nowiki>#</nowiki>27, <nowiki>#</nowiki>3-<nowiki>#</nowiki>29, <nowiki>#</nowiki>3-23-<nowiki>#</nowiki>5, <nowiki>#</nowiki>33-pSB1AK3, <nowiki>#</nowiki>34--pSB1AK3, <nowiki>#</nowiki>35--pSB1AK3)
+
*Ligation(<nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">24</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">24</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-</html><nowiki>#</nowiki><html><span class="part_num">28</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">27</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">29</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">23</span>-</html><nowiki>#</nowiki><html><span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">33</span>-</html>pSB1AK3, <nowiki>#</nowiki><html><span class="part_num">34</span>--</html>pSB1AK3, <nowiki>#</nowiki><html><span class="part_num">35</span>--</html>pSB1AK3)
-
*Transformation(ligation products:WT, <nowiki>#</nowiki>2-9:cheZ-, <nowiki>#</nowiki>14, <nowiki>#</nowiki>17) -> 37 incubation
+
*Transformation(ligation products:WT, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">9</span></html>:cheZ-, <nowiki>#</nowiki><html><span class="part_num">14</span></html>, <nowiki>#</nowiki><html><span class="part_num">17</span></html>) -> 37 incubation
<html></div></html>
<html></div></html>
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/08/16(Tue)===
==='11/08/16(Tue)===
-
*Colony PCR(<nowiki>#</nowiki>3-<nowiki>#</nowiki>14-5, <nowiki>#</nowiki>24-<nowiki>#</nowiki>3-17-5, <nowiki>#</nowiki>20-<nowiki>#</nowiki>28, <nowiki>#</nowiki>3-<nowiki>#</nowiki>27, <nowiki>#</nowiki>3-<nowiki>#</nowiki>29, <nowiki>#</nowiki>3-23-<nowiki>#</nowiki>5)
+
*Colony PCR(<nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">24</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-</html><nowiki>#</nowiki><html><span class="part_num">28</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">27</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">29</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">23</span>-</html><nowiki>#</nowiki><html><span class="part_num">5</span></html>)
-
*Culture (<nowiki>#</nowiki>20-<nowiki>#</nowiki>28, <nowiki>#</nowiki>5, <nowiki>#</nowiki>14, <nowiki>#</nowiki>17, <nowiki>#</nowiki>2-9, <nowiki>#</nowiki>2-4, <nowiki>#</nowiki>2-3-17-5, <nowiki>#</nowiki>1, <nowiki>#</nowiki>2)
+
*Culture (<nowiki>#</nowiki><html><span class="part_num">20</span>-</html><nowiki>#</nowiki><html><span class="part_num">28</span></html>, <nowiki>#</nowiki><html><span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">14</span></html>, <nowiki>#</nowiki><html><span class="part_num">17</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">9</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">4</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">1</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span></html>)
-
*Incubation (<nowiki>#</nowiki>24-<nowiki>#</nowiki>3-11-5, <nowiki>#</nowiki>33-pSB1AK3, <nowiki>#</nowiki>34-pSB1AK3, <nowiki>#</nowiki>35-pSB1AK3)
+
*Incubation (<nowiki>#</nowiki><html><span class="part_num">24</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">33</span>-</html>pSB1AK3, <nowiki>#</nowiki><html><span class="part_num">34</span>-</html>pSB1AK3, <nowiki>#</nowiki><html><span class="part_num">35</span>-</html>pSB1AK3)
*Colony PCR(additional)(previously incubated ligation products: 3, 8, 10)
*Colony PCR(additional)(previously incubated ligation products: 3, 8, 10)
*hotstart PCR(sulAp, uvrAp, uvrBp)
*hotstart PCR(sulAp, uvrAp, uvrBp)
-
*Digest(SPcut: <nowiki>#</nowiki>3, Ecut:<nowiki>#</nowiki>3(control)) go to 37 incubation
+
*Digest(SPcut: <nowiki>#</nowiki><html><span class="part_num">3</span></html>, Ecut:<nowiki>#</nowiki><html><span class="part_num">3</span></html>(control)) go to 37 incubation
-
*Ligation retry(<nowiki>#</nowiki>3-<nowiki>#</nowiki>14-5, <nowiki>#</nowiki>3-<nowiki>#</nowiki>27, <nowiki>#</nowiki>3-<nowiki>#</nowiki>29, Negative control :<nowiki>#</nowiki>3)
+
*Ligation retry(<nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">27</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">29</span></html>, Negative control :<nowiki>#</nowiki><html><span class="part_num">3</span></html>)
-
*Frozen stock(<nowiki>#</nowiki>14, <nowiki>#</nowiki>17)
+
*Frozen stock(<nowiki>#</nowiki><html><span class="part_num">14</span></html>, <nowiki>#</nowiki><html><span class="part_num">17</span></html>)
-
*Culture test(<nowiki>#</nowiki>14 on 0.3%Agar.(km))
+
*Culture test(<nowiki>#</nowiki><html><span class="part_num">14</span></html> on 0.3%Agar.(km))
*Ethanol precipitation: 29.9ng/ul * 1000ul
*Ethanol precipitation: 29.9ng/ul * 1000ul
-
*Miniprep(Ligation products : <nowiki>#</nowiki>2-3-17-5, <nowiki>#</nowiki>14, <nowiki>#</nowiki>17, <nowiki>#</nowiki>1, <nowiki>#</nowiki>2)
+
*Miniprep(Ligation products : <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">14</span></html>, <nowiki>#</nowiki><html><span class="part_num">17</span></html>, <nowiki>#</nowiki><html><span class="part_num">1</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span></html>)
*Electrophoresis(Colony PCR products: 3, 8, 10)
*Electrophoresis(Colony PCR products: 3, 8, 10)
-
*Waking from frozen stock(<nowiki>#</nowiki>3, CheZ-, WT)
+
*Waking from frozen stock(<nowiki>#</nowiki><html><span class="part_num">3</span></html>, CheZ-, WT)
<html></div></html>
<html></div></html>
Line 580: Line 583:
*Making Competent Cell, SOB medium
*Making Competent Cell, SOB medium
*Ethanol precipitation(WT: 40ng/ul)
*Ethanol precipitation(WT: 40ng/ul)
-
*Culture from master-plate(<nowiki>#</nowiki>6-5, <nowiki>#</nowiki>20-28, <nowiki>#</nowiki>3-27, <nowiki>#</nowiki>33, <nowiki>#</nowiki>24-3-11-5 <nowiki>#</nowiki>24-3-17-5) -> Miniprep
+
*Culture from master-plate(<nowiki>#</nowiki><html><span class="part_num">6</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">28</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span></html>, <nowiki>#</nowiki><html><span class="part_num">33</span></html>, <nowiki>#</nowiki><html><span class="part_num">24</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html> <nowiki>#</nowiki><html><span class="part_num">24</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>) -> Miniprep
*Assay(IPTG induced cell lysis device
*Assay(IPTG induced cell lysis device
-
*Gel extraction(SPcut:<nowiki>#</nowiki>3)
+
*Gel extraction(SPcut:<nowiki>#</nowiki><html><span class="part_num">3</span></html>)
-
*Culture from Frozen stock(<nowiki>#</nowiki>1, <nowiki>#</nowiki>2, <nowiki>#</nowiki>2-3-17-5)
+
*Culture from Frozen stock(<nowiki>#</nowiki><html><span class="part_num">1</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
*Electrophresis check(Loading only DNA size markers)
*Electrophresis check(Loading only DNA size markers)
-
*Miniprep(<nowiki>#</nowiki>3, <nowiki>#</nowiki>6-5, <nowiki>#</nowiki>20-28, <nowiki>#</nowiki>3-27, <nowiki>#</nowiki>33, <nowiki>#</nowiki>24-3-11-5 <nowiki>#</nowiki>24-3-17-5)
+
*Miniprep(<nowiki>#</nowiki><html><span class="part_num">3</span></html>, <nowiki>#</nowiki><html><span class="part_num">6</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">28</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span></html>, <nowiki>#</nowiki><html><span class="part_num">33</span></html>, <nowiki>#</nowiki><html><span class="part_num">24</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html> <nowiki>#</nowiki><html><span class="part_num">24</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
<html></div></html>
<html></div></html>
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/08/18(Thu)===
==='11/08/18(Thu)===
-
*Digest(8/17<nowiki>#</nowiki>3, <nowiki>#</nowiki>33 SPcut, <nowiki>#</nowiki>14-5 XPcut, <nowiki>#</nowiki>33 Ecut, <nowiki>#</nowiki>3-27,<nowiki>#</nowiki>3-30,<nowiki>#</nowiki>20-28 EScut, pSB1K3 EPcut)
+
*Digest(8/17<nowiki>#</nowiki><html><span class="part_num">3</span></html>, <nowiki>#</nowiki><html><span class="part_num">33</span></html> SPcut, <nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html> XPcut, <nowiki>#</nowiki><html><span class="part_num">33</span></html> Ecut, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span></html>,<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">30</span></html>,<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">28</span></html> EScut, pSB1K3 EPcut)
*Hotstart-PCR (Change conditions a little)
*Hotstart-PCR (Change conditions a little)
**From Genome :  uvrBp, cheZ
**From Genome :  uvrBp, cheZ
**From nested-region : uvrA
**From nested-region : uvrA
-
*Culture from 0809 master-plate ⑪ (<nowiki>#</nowiki>2-3-11-5) → Miniprep.
+
*Culture from 0809 master-plate ⑪ (<nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>) → Miniprep.
*Making LB plate & tube with various agarose conc. (plate: 0.5, 0.7, 0.9%×2 each, tube: 0.5,1.0,1.5,2.0,3.0%×2 each)(selection: Kanamycin)
*Making LB plate & tube with various agarose conc. (plate: 0.5, 0.7, 0.9%×2 each, tube: 0.5,1.0,1.5,2.0,3.0%×2 each)(selection: Kanamycin)
-
*Gel extraction(8/17<nowiki>#</nowiki>3_SP,<nowiki>#</nowiki>33_SP, <nowiki>#</nowiki>14-5_XP, <nowiki>#</nowiki>33_E, <nowiki>#</nowiki>3-27_ES, pSB1K3 _EP)
+
*Gel extraction(8/17<nowiki>#</nowiki><html><span class="part_num">3</span></html>_SP,<nowiki>#</nowiki><html><span class="part_num">33</span></html>_SP, <nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">33</span></html>_E, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span></html>_ES, pSB1K3 _EP)
-
*Miniprep(<nowiki>#</nowiki>1, <nowiki>#</nowiki>2, <nowiki>#</nowiki>2-3-11-5, <nowiki>#</nowiki>2-3-17-5)
+
*Miniprep(<nowiki>#</nowiki><html><span class="part_num">1</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
-
*Digest(<nowiki>#</nowiki>1,<nowiki>#</nowiki>2 SPcut, <nowiki>#</nowiki>2-3-17-5 XPcut, <nowiki>#</nowiki>2-3-11-5 SPcut(check), EPcut(selection), <nowiki>#</nowiki>3-30, <nowiki>#</nowiki>20-28 EScut, 0817 Gel extracted PCR product (uvrBp) EScut, 0818PCR products (uvrAp, uvrBp, cheZ) ES&XPcut)
+
*Digest(<nowiki>#</nowiki><html><span class="part_num">1</span></html>,<nowiki>#</nowiki><html><span class="part_num">2</span></html> SPcut, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html> XPcut, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html> SPcut(check), EPcut(selection), <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">30</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">28</span></html> EScut, 0817 Gel extracted PCR product (uvrBp) EScut, 0818PCR products (uvrAp, uvrBp, cheZ) ES&XPcut)
-
*Ligation (8/17<nowiki>#</nowiki>3-<nowiki>#</nowiki>14-5, 8/17<nowiki>#</nowiki>3-8/15<nowiki>#</nowiki>29, <nowiki>#</nowiki>33-<nowiki>#</nowiki>3-11-5, 8/15<nowiki>#</nowiki>34-pSB1AK3, <nowiki>#</nowiki>3-27-<nowiki>#</nowiki>5, <nowiki>#</nowiki>20-28-pSB1AK3, <nowiki>#</nowiki>28-pSB1AK3)
+
*Ligation (8/17<nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>, 8/17<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">8</span></html>/15<nowiki>#</nowiki><html><span class="part_num">29</span></html>, <nowiki>#</nowiki><html><span class="part_num">33</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>, 8/15<nowiki>#</nowiki><html><span class="part_num">34</span>-</html>pSB1AK3, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-</html><nowiki>#</nowiki><html><span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">28</span>-</html>pSB1AK3, <nowiki>#</nowiki><html><span class="part_num">28</span>-</html>pSB1AK3)
*Culture for frozen stock in eppendolf tube
*Culture for frozen stock in eppendolf tube
<html></div></html>
<html></div></html>
Line 605: Line 608:
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/08/19(Fri)===
==='11/08/19(Fri)===
-
*ColonyPCR(from ligation products)(<nowiki>#</nowiki>3-14-5, <nowiki>#</nowiki>3-29, <nowiki>#</nowiki>33-3-11-5, <nowiki>#</nowiki>34-pSB1AK3, <nowiki>#</nowiki>3-27-5)
+
*ColonyPCR(from ligation products)(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span></html>, <nowiki>#</nowiki><html><span class="part_num">33</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">34</span>-</html>pSB1AK3, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>)
-
*Gel extraction(08/18's digest products:<nowiki>#</nowiki>1_SP, <nowiki>#</nowiki>2_SP, <nowiki>#</nowiki>2-3-11-5_SP, <nowiki>#</nowiki>2-3-17-5_XP, cheZ_XP, <nowiki>#</nowiki>2-3-11-5_EP, 8/18uvrBp_ES, 8/17uvrBp_ES, uvrAp_ES)
+
*Gel extraction(08/18's digest products:<nowiki>#</nowiki><html><span class="part_num">1</span></html>_SP, <nowiki>#</nowiki><html><span class="part_num">2</span></html>_SP, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>_SP, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_XP, cheZ_XP, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>_EP, 8/18uvrBp_ES, 8/17uvrBp_ES, uvrAp_ES)
-
*Colony PCR retry(<nowiki>#</nowiki>3-29, <nowiki>#</nowiki>34-pSB1AK3)
+
*Colony PCR retry(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span></html>, <nowiki>#</nowiki><html><span class="part_num">34</span>-</html>pSB1AK3)
*Frozen Stock
*Frozen Stock
*Check the list of frozen stock
*Check the list of frozen stock
*Assay of recAp from biobrick parts using RFP construct & transilluminator
*Assay of recAp from biobrick parts using RFP construct & transilluminator
-
*Electrophoresis(<nowiki>#</nowiki>3, <nowiki>#</nowiki>3-27, <nowiki>#</nowiki>3-30, <nowiki>#</nowiki>6-5, <nowiki>#</nowiki>7, <nowiki>#</nowiki>20-28, <nowiki>#</nowiki>24-3-11-5, <nowiki>#</nowiki>24-3-17-5, <nowiki>#</nowiki>33)
+
*Electrophoresis(<nowiki>#</nowiki><html><span class="part_num">3</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">30</span></html>, <nowiki>#</nowiki><html><span class="part_num">6</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">7</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">28</span></html>, <nowiki>#</nowiki><html><span class="part_num">24</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">24</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">33</span></html>)
<html></div></html>
<html></div></html>
Line 618: Line 621:
*Making SOB medium
*Making SOB medium
*Culture for Miniprep
*Culture for Miniprep
-
**From frozen stock(<nowiki>#</nowiki>1, <nowiki>#</nowiki>2, <nowiki>#</nowiki>7, <nowiki>#</nowiki>2-3-17-5(2), <nowiki>#</nowiki>3-30)
+
**From frozen stock(<nowiki>#</nowiki><html><span class="part_num">1</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span></html>, <nowiki>#</nowiki><html><span class="part_num">7</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>(2), <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">30</span></html>)
-
**From master plate(<nowiki>#</nowiki>3-14-5, <nowiki>#</nowiki>33-3-11-5, <nowiki>#</nowiki>34-pSB1AK3, <nowiki>#</nowiki>3-27-5)
+
**From master plate(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">33</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">34</span>-</html>pSB1AK3, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>)
**From master plate retry( at 13:50 OD600=0)
**From master plate retry( at 13:50 OD600=0)
**From ligation products (for mistake above)
**From ligation products (for mistake above)
*Miniprep, Nanodrop and Electrophoresis
*Miniprep, Nanodrop and Electrophoresis
-
**From frozen stock(<nowiki>#</nowiki>1, <nowiki>#</nowiki>2, <nowiki>#</nowiki>2-3-17-5(2), <nowiki>#</nowiki>3-30) <nowiki>#</nowiki>7 was a little bit weak
+
**From frozen stock(<nowiki>#</nowiki><html><span class="part_num">1</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>(2), <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">30</span></html>) <nowiki>#</nowiki><html><span class="part_num">7</span></html> was a little bit weak
-
**From ligation products(<nowiki>#</nowiki>7, <nowiki>#</nowiki>3-14-5, <nowiki>#</nowiki>3-27-5, <nowiki>#</nowiki>34, <nowiki>#</nowiki>33-3-11-5)
+
**From ligation products(<nowiki>#</nowiki><html><span class="part_num">7</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">34</span></html>, <nowiki>#</nowiki><html><span class="part_num">33</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>)
*Digest
*Digest
-
**From Frozen Stock (<nowiki>#</nowiki>7_EX, <nowiki>#</nowiki>30_XP, <nowiki>#</nowiki>2-3-17-5_XP, <nowiki>#</nowiki>3-30_ES)
+
**From Frozen Stock (<nowiki>#</nowiki><html><span class="part_num">7</span></html>_EX, <nowiki>#</nowiki><html><span class="part_num">30</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">30</span></html>_ES)
-
**From Master plate(<nowiki>#</nowiki>3-14-5_XP, <nowiki>#</nowiki>34-pSB1AK3_SP, <nowiki>#</nowiki>3-27-5_XP)
+
**From Master plate(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">34</span>-</html>pSB1AK3_SP, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>_XP)
-
*Sulvage <nowiki>#</nowiki>33 using protocol of transformation  
+
*Sulvage <nowiki>#</nowiki><html><span class="part_num">33</span></html> using protocol of transformation  
*Culture for Frozen stock
*Culture for Frozen stock
-
**From Master plate(<nowiki>#</nowiki>24-3-17-5, <nowiki>#</nowiki>14-5), 700ul, in eppendolf tube
+
**From Master plate(<nowiki>#</nowiki><html><span class="part_num">24</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>), 700ul, in eppendolf tube
<html></div></html>
<html></div></html>
Line 636: Line 639:
==='11/08/23(Tue)===
==='11/08/23(Tue)===
*Gel extraction
*Gel extraction
-
**from O.N. digest products(<nowiki>#</nowiki>7_EX, <nowiki>#</nowiki>30_XP, <nowiki>#</nowiki>2-3-17-5_XP, <nowiki>#</nowiki>3-30_ES, <nowiki>#</nowiki>3-14-5_XP, <nowiki>#</nowiki>34-pSB1AK3_SP, <nowiki>#</nowiki>3-27-5_XP)
+
**from O.N. digest products(<nowiki>#</nowiki><html><span class="part_num">7</span></html>_EX, <nowiki>#</nowiki><html><span class="part_num">30</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">30</span></html>_ES, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">34</span>-</html>pSB1AK3_SP, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>_XP)
*Frozen stock
*Frozen stock
-
**From O.N. culture in eppendolf tube 700ul(<nowiki>#</nowiki>24-3-17-5, <nowiki>#</nowiki>14-5)
+
**From O.N. culture in eppendolf tube 700ul(<nowiki>#</nowiki><html><span class="part_num">24</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>)
-
**From O.N. culture(for Miniprep)(<nowiki>#</nowiki>34, <nowiki>#</nowiki>3-27-5, <nowiki>#</nowiki>33-3-11-5)
+
**From O.N. culture(for Miniprep)(<nowiki>#</nowiki><html><span class="part_num">34</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">33</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>)
*Miniprep
*Miniprep
-
**retry from master plate <nowiki>#</nowiki>3-14-5(278.1 ng/ul), <nowiki>#</nowiki>33-3-11-5(188.9ng/ul), <nowiki>#</nowiki>34-pSB1AK3(89.6ng/ul), <nowiki>#</nowiki>3-27-5(327.0 ng/ul) O.N. culture
+
**retry from master plate <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>(278.1 ng/ul), <nowiki>#</nowiki><html><span class="part_num">33</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>(188.9ng/ul), <nowiki>#</nowiki><html><span class="part_num">34</span>-</html>pSB1AK3(89.6ng/ul), <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>(327.0 ng/ul) O.N. culture
-
*Digest (<nowiki>#</nowiki>14-5 XPcut, <nowiki>#</nowiki>3-27-5 Xcut, Pcut (check), <nowiki>#</nowiki>3-27-5 EScut (sulvage) )
+
*Digest (<nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html> XPcut, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html> Xcut, Pcut (check), <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html> EScut (sulvage) )
-
*Ligation (<nowiki>#</nowiki>14-<nowiki>#</nowiki>5, <nowiki>#</nowiki>3-30-<nowiki>#</nowiki>5, <nowiki>#</nowiki>3-<nowiki>#</nowiki>30, <nowiki>#</nowiki>34-<nowiki>#</nowiki>3-11-5 <nowiki>#</nowiki>20-<nowiki>#</nowiki>28)
+
*Ligation (<nowiki>#</nowiki><html><span class="part_num">14</span>-</html><nowiki>#</nowiki><html><span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">30</span>-</html><nowiki>#</nowiki><html><span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">30</span></html>, <nowiki>#</nowiki><html><span class="part_num">34</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html> <nowiki>#</nowiki><html><span class="part_num">20</span>-</html><nowiki>#</nowiki><html><span class="part_num">28</span></html>)
-
*<nowiki>#</nowiki>33 sulvage (isolation culture)
+
*<nowiki>#</nowiki><html><span class="part_num">33</span></html> sulvage (isolation culture)
*Making culture(LB amp, LB)
*Making culture(LB amp, LB)
<html></div></html>
<html></div></html>
Line 650: Line 653:
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/08/24(Wed)===
==='11/08/24(Wed)===
-
*Gel extraction & Electrophoresis(<nowiki>#</nowiki>3-27-5_ES, <nowiki>#</nowiki>14-5_XP, <nowiki>#</nowiki>3-27-5_X, *PCR for amplification (or cloning) -> PCR clean-up system -> digest
+
*Gel extraction & Electrophoresis(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>_ES, <nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>_X, *PCR for amplification (or cloning) -> PCR clean-up system -> digest
-
**<nowiki>#</nowiki>27 (82.1 ng/ul, total = 30 ul) -> XP cut
+
**<nowiki>#</nowiki><html><span class="part_num">27</span></html> (82.1 ng/ul, total = 30 ul) -> XP cut
-
**<nowiki>#</nowiki>28 (104.4 ng/ul, total = 30 ul) -> XP cut
+
**<nowiki>#</nowiki><html><span class="part_num">28</span></html> (104.4 ng/ul, total = 30 ul) -> XP cut
*Making plate and amp stock ->4℃
*Making plate and amp stock ->4℃
**amp 32
**amp 32
**km 11 (km conc. = 3/4 * that it was)
**km 11 (km conc. = 3/4 * that it was)
*Miniprep and Nanodrop
*Miniprep and Nanodrop
-
**<nowiki>#</nowiki>33 32.1 ng/ul <- there is probability that culture volume was not enough ->transfromation
+
**<nowiki>#</nowiki><html><span class="part_num">33</span></html> 32.1 ng/ul <- there is probability that culture volume was not enough ->transfromation
*Luciferase assay reagents prep
*Luciferase assay reagents prep
**D-Luciferin Solution : 1ml*10 + 100ul*10 at -80℃
**D-Luciferin Solution : 1ml*10 + 100ul*10 at -80℃
**Coelenterazine Solution : 100ul * 10 at -20℃
**Coelenterazine Solution : 100ul * 10 at -20℃
**Lysis buffer : 500ul * 10 at -20℃
**Lysis buffer : 500ul * 10 at -20℃
-
*Ligation(<nowiki>#</nowiki>3-30-<nowiki>#</nowiki>5, <nowiki>#</nowiki>3-11-5-<nowiki>#</nowiki>34, <nowiki>#</nowiki>30-<nowiki>#</nowiki>3, <nowiki>#</nowiki>28-<nowiki>#</nowiki>20, <nowiki>#</nowiki>35-pSB1AK3, <nowiki>#</nowiki>2-3-11-5-pSB1K3, <nowiki>#</nowiki>14-5-<nowiki>#</nowiki>3 and Negative control)
+
*Ligation(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">30</span>-</html><nowiki>#</nowiki><html><span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span>-</html><nowiki>#</nowiki><html><span class="part_num">34</span></html>, <nowiki>#</nowiki><html><span class="part_num">30</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span></html>, <nowiki>#</nowiki><html><span class="part_num">28</span>-</html><nowiki>#</nowiki><html><span class="part_num">20</span></html>, <nowiki>#</nowiki><html><span class="part_num">35</span>-</html>pSB1AK3, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span>-</html>pSB1K3, <nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span></html> and Negative control)
<html></div></html>
<html></div></html>
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/08/25(Thu)===
==='11/08/25(Thu)===
-
*Renilla luciferase assay (with <nowiki>#</nowiki>2-3-17-5 culture)
+
*Renilla luciferase assay (with <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html> culture)
-
*Gel extraction (<nowiki>#</nowiki>27, <nowiki>#</nowiki>28 XPcut)
+
*Gel extraction (<nowiki>#</nowiki><html><span class="part_num">27</span></html>, <nowiki>#</nowiki><html><span class="part_num">28</span></html> XPcut)
-
*Ligation (0824 retry & <nowiki>#</nowiki>20-<nowiki>#</nowiki>28, <nowiki>#</nowiki>3-<nowiki>#</nowiki>27)
+
*Ligation (0824 retry & <nowiki>#</nowiki><html><span class="part_num">20</span>-</html><nowiki>#</nowiki><html><span class="part_num">28</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">27</span></html>)
-
*Transformation (Ligation products into WT competent cells & <nowiki>#</nowiki>14-5 miniprep product into WT cell (line-drawing spread), <nowiki>#</nowiki>33 miniprep product into ΔcheZ cell (line-drawing spread))  
+
*Transformation (Ligation products into WT competent cells & <nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html> miniprep product into WT cell (line-drawing spread), <nowiki>#</nowiki><html><span class="part_num">33</span></html> miniprep product into ΔcheZ cell (line-drawing spread))  
-
*Digest (<nowiki>#</nowiki>14 XPcut)
+
*Digest (<nowiki>#</nowiki><html><span class="part_num">14</span></html> XPcut)
-
*Passage(<nowiki>#</nowiki>2-3-17-5, WT, ΔcheZ, WT in eppen, ΔcheZ in eppen)
+
*Passage(<nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, WT, ΔcheZ, WT in eppen, ΔcheZ in eppen)
<html></div></html>
<html></div></html>
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/08/26(Fri)===
==='11/08/26(Fri)===
-
*Renilla luciferase assay (with <nowiki>#</nowiki>2-3-17-5 culture)
+
*Renilla luciferase assay (with <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html> culture)
-
*Colony PCR (<nowiki>#</nowiki>33 Miniprep transformations)
+
*Colony PCR (<nowiki>#</nowiki><html><span class="part_num">33</span></html> Miniprep transformations)
-
*Gel extraction (<nowiki>#</nowiki>14_XP)
+
*Gel extraction (<nowiki>#</nowiki><html><span class="part_num">14</span></html>_XP)
*Ligation (retry) (using today's competent cells, 0802 ΔcheZ cells)
*Ligation (retry) (using today's competent cells, 0802 ΔcheZ cells)
*Cloning (cheZ)
*Cloning (cheZ)
-
*Digest (<nowiki>#</nowiki>5 EXcut, cheZ PCR products XPcut)
+
*Digest (<nowiki>#</nowiki><html><span class="part_num">5</span></html> EXcut, cheZ PCR products XPcut)
-
*Passage in a shaker(<nowiki>#</nowiki>2-3-17-5, WT, ΔcheZ)
+
*Passage in a shaker(<nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, WT, ΔcheZ)
*Making competent cell, LB broth
*Making competent cell, LB broth
*Contamination check (0817 comp., 0802 comp., 0802 ⊿cheZ comp. spread on LB (amp) plates)
*Contamination check (0817 comp., 0802 comp., 0802 ⊿cheZ comp. spread on LB (amp) plates)
Line 699: Line 702:
*Making ΔcheZ comp., LB amp plates, TB
*Making ΔcheZ comp., LB amp plates, TB
*Colony PCR & master plate preparation -> Failure!!
*Colony PCR & master plate preparation -> Failure!!
-
*Gel extraction (<nowiki>#</nowiki>29_XP, <nowiki>#</nowiki>5_EX)
+
*Gel extraction (<nowiki>#</nowiki><html><span class="part_num">29</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">5</span></html>_EX)
-
*Miniprep and frozen stocks (<nowiki>#</nowiki>33, <nowiki>#</nowiki>14-5)
+
*Miniprep and frozen stocks (<nowiki>#</nowiki><html><span class="part_num">33</span></html>, <nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>)
-
*Digest (<nowiki>#</nowiki>20 SPcut, <nowiki>#</nowiki>14-5 XPcut)
+
*Digest (<nowiki>#</nowiki><html><span class="part_num">20</span></html> SPcut, <nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html> XPcut)
-
*Waking from frozen stock (WT, ⊿cheZ, <nowiki>#</nowiki>20, <nowiki>#</nowiki>14-5)
+
*Waking from frozen stock (WT, ⊿cheZ, <nowiki>#</nowiki><html><span class="part_num">20</span></html>, <nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>)
<html></div></html>
<html></div></html>
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/08/29(Mon)===
==='11/08/29(Mon)===
-
*PCR purification -> digest with XP(<nowiki>#</nowiki>27, <nowiki>#</nowiki>28)
+
*PCR purification -> digest with XP(<nowiki>#</nowiki><html><span class="part_num">27</span></html>, <nowiki>#</nowiki><html><span class="part_num">28</span></html>)
-
*Miniprep(<nowiki>#</nowiki>20, <nowiki>#</nowiki>14-5)
+
*Miniprep(<nowiki>#</nowiki><html><span class="part_num">20</span></html>, <nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>)
-
*Gel extraction(<nowiki>#</nowiki>14-5, <nowiki>#</nowiki>20)
+
*Gel extraction(<nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span></html>)
-
*Ligation (higher I/V ratio, into cheZ comp.)(<nowiki>#</nowiki>3-<nowiki>#</nowiki>14-5(retry), <nowiki>#</nowiki>2-3-11-5 to pSB1K3 (retry), <nowiki>#</nowiki>35-pSB1AK3, (<nowiki>#</nowiki>33,<nowiki>#</nowiki>34)-<nowiki>#</nowiki>3-17-5)
+
*Ligation (higher I/V ratio, into cheZ comp.)(<nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>(retry), <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html> to pSB1K3 (retry), <nowiki>#</nowiki><html><span class="part_num">35</span>-</html>pSB1AK3, (<nowiki>#</nowiki><html><span class="part_num">33</span></html>,<nowiki>#</nowiki><html><span class="part_num">34</span></html>)-<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
<html></div></html>
<html></div></html>
Line 716: Line 719:
==='11/08/30(Tue)===
==='11/08/30(Tue)===
*Making LB amp, Aspartic acid solution (10mM), H2O2 solution (10mM)
*Making LB amp, Aspartic acid solution (10mM), H2O2 solution (10mM)
-
*Gel extraction -> PCR was failed!(<nowiki>#</nowiki>27,<nowiki>#</nowiki>28)
+
*Gel extraction -> PCR was failed!(<nowiki>#</nowiki><html><span class="part_num">27</span></html>,<nowiki>#</nowiki><html><span class="part_num">28</span></html>)
*There was trash (from PCR) below
*There was trash (from PCR) below
-
*Colony PCR(<nowiki>#</nowiki>3-<nowiki>#</nowiki>14-5, <nowiki>#</nowiki>2-3-11-5 to pSB1K3, <nowiki>#</nowiki>35-pSB1AK3, (<nowiki>#</nowiki>33,<nowiki>#</nowiki>34)-<nowiki>#</nowiki>3-17-5)
+
*Colony PCR(<nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html> to pSB1K3, <nowiki>#</nowiki><html><span class="part_num">35</span>-</html>pSB1AK3, (<nowiki>#</nowiki><html><span class="part_num">33</span></html>,<nowiki>#</nowiki><html><span class="part_num">34</span></html>)-<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
-
*PCR -> check (5uL EP -> PCR clean-up(<nowiki>#</nowiki>27,<nowiki>#</nowiki>28)
+
*PCR -> check (5uL EP -> PCR clean-up(<nowiki>#</nowiki><html><span class="part_num">27</span></html>,<nowiki>#</nowiki><html><span class="part_num">28</span></html>)
-
*Digest(<nowiki>#</nowiki>20_SP, pSB1AK3_EP, <nowiki>#</nowiki>3_SP)
+
*Digest(<nowiki>#</nowiki><html><span class="part_num">20</span></html>_SP, pSB1AK3_EP, <nowiki>#</nowiki><html><span class="part_num">3</span></html>_SP)
-
*EP for MP check(<nowiki>#</nowiki>14-5, <nowiki>#</nowiki>20)
+
*EP for MP check(<nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span></html>)
-
*Ligation(<nowiki>#</nowiki>3-<nowiki>#</nowiki>29, pSB1AK3-<nowiki>#</nowiki>2-3-11-5 on Amp/Km plate, <nowiki>#</nowiki>3 (Negative control), pSB1AK3 (Negative control) on Amp/Km plate
+
*Ligation(<nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">29</span></html>, pSB1AK3-<nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html> on Amp/Km plate, <nowiki>#</nowiki><html><span class="part_num">3</span></html> (Negative control), pSB1AK3 (Negative control) on Amp/Km plate
-
*Transformation(<nowiki>#</nowiki>2-9 into cheZ-) -> Line-drawing plating
+
*Transformation(<nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">9</span></html> into cheZ-) -> Line-drawing plating
-
*SOS stress (UV irradiation or H2O2)(<nowiki>#</nowiki>24-3-11-5, <nowiki>#</nowiki>33-3-11-5)
+
*SOS stress (UV irradiation or H2O2)(<nowiki>#</nowiki><html><span class="part_num">24</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">33</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>)
-
*culture for Miniprep from master plate(<nowiki>#</nowiki>34-3-17-5)
+
*culture for Miniprep from master plate(<nowiki>#</nowiki><html><span class="part_num">34</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
-
*Waking from frozen stock(<nowiki>#</nowiki>24-3-11-5, <nowiki>#</nowiki>33-3-11-5)
+
*Waking from frozen stock(<nowiki>#</nowiki><html><span class="part_num">24</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">33</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>)
<html></div></html>
<html></div></html>
Line 734: Line 737:
*Luciferin reagent preparation
*Luciferin reagent preparation
*Flask A.C. for making competent cells
*Flask A.C. for making competent cells
-
*Miniprep (<nowiki>#</nowiki>34-3-17-5)
+
*Miniprep (<nowiki>#</nowiki><html><span class="part_num">34</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
-
*Gel extraction(<nowiki>#</nowiki>20_SP, pSB1AK3_EP, <nowiki>#</nowiki>3_SP)
+
*Gel extraction(<nowiki>#</nowiki><html><span class="part_num">20</span></html>_SP, pSB1AK3_EP, <nowiki>#</nowiki><html><span class="part_num">3</span></html>_SP)
-
*PCR -> clean-up -> XPcut(<nowiki>#</nowiki>27, <nowiki>#</nowiki>28, <nowiki>#</nowiki>14-5, <nowiki>#</nowiki>3-30)
+
*PCR -> clean-up -> XPcut(<nowiki>#</nowiki><html><span class="part_num">27</span></html>, <nowiki>#</nowiki><html><span class="part_num">28</span></html>, <nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">30</span></html>)
-
*Waking from frozen stock(<nowiki>#</nowiki>2-9 (WT), <nowiki>#</nowiki>24-3-17-5)
+
*Waking from frozen stock(<nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">9</span></html> (WT), <nowiki>#</nowiki><html><span class="part_num">24</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
-
*Plating from liquid culture(<nowiki>#</nowiki>2-3-17-5)
+
*Plating from liquid culture(<nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
-
*Ligation(<nowiki>#</nowiki>20-<nowiki>#</nowiki>3-17-5, <nowiki>#</nowiki>2-3-11-5-pSB1AK3, <nowiki>#</nowiki>3-<nowiki>#</nowiki>14, <nowiki>#</nowiki>20-<nowiki>#</nowiki>28, <nowiki>#</nowiki>3-<nowiki>#</nowiki>27, <nowiki>#</nowiki>3-<nowiki>#</nowiki>29, <nowiki>#</nowiki>35-pSB1AK3, <nowiki>#</nowiki>3-<nowiki>#</nowiki>14-5, <nowiki>#</nowiki>3-30-<nowiki>#</nowiki>5, <nowiki>#</nowiki>33-<nowiki>#</nowiki>3-17-5 & Negative control)
+
*Ligation(<nowiki>#</nowiki><html><span class="part_num">20</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span>-</html>pSB1AK3, <nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">14</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-</html><nowiki>#</nowiki><html><span class="part_num">28</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">27</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">29</span></html>, <nowiki>#</nowiki><html><span class="part_num">35</span>-</html>pSB1AK3, <nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">30</span>-</html><nowiki>#</nowiki><html><span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">33</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html> & Negative control)
*Transformation
*Transformation
-
**Plasmid isolation(<nowiki>#</nowiki>34-3-17-5(MP;1:9gradient))
+
**Plasmid isolation(<nowiki>#</nowiki><html><span class="part_num">34</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>(MP;1:9gradient))
-
**Part cloning(<nowiki>#</nowiki>36, <nowiki>#</nowiki>37)
+
**Part cloning(<nowiki>#</nowiki><html><span class="part_num">36</span></html>, <nowiki>#</nowiki><html><span class="part_num">37</span></html>)
**Contamination check(pSB1AK3(EPcut), pSB1K3(EPcut))
**Contamination check(pSB1AK3(EPcut), pSB1K3(EPcut))
-
**Titer check(<nowiki>#</nowiki>3-17-5(Takara comp.,1:9gradient), <nowiki>#</nowiki>2-3-11-5(ΔcheZ comp.0802, 1:9gradient), <nowiki>#</nowiki>2-3-11-5(ΔcheZ comp.0830, 1:9gradient))
+
**Titer check(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>(Takara comp.,1:9gradient), <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>(ΔcheZ comp.0802, 1:9gradient), <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>(ΔcheZ comp.0830, 1:9gradient))
**Ligation products
**Ligation products
<html></div></html>
<html></div></html>
Line 754: Line 757:
==='11/09/01(Thu)===
==='11/09/01(Thu)===
*Colony PCR
*Colony PCR
-
**<nowiki>#</nowiki>34-3-17-5(MP), <nowiki>#</nowiki>3-14, <nowiki>#</nowiki>3-29, <nowiki>#</nowiki>35-pSB1AK3, <nowiki>#</nowiki>3-14-5, <nowiki>#</nowiki>37(from Part cloning)
+
**<nowiki>#</nowiki><html><span class="part_num">34</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>(MP), <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span></html>, <nowiki>#</nowiki><html><span class="part_num">35</span>-</html>pSB1AK3, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">37</span></html>(from Part cloning)
*Gel extraction
*Gel extraction
-
**<nowiki>#</nowiki>27_XP ,<nowiki>#</nowiki>28_XP(0830); <nowiki>#</nowiki>27_XP, <nowiki>#</nowiki>28_XP, <nowiki>#</nowiki>3-30_XP -> failed!, <nowiki>#</nowiki>14-5_XP(0831)
+
**<nowiki>#</nowiki><html><span class="part_num">27</span></html>_XP ,<nowiki>#</nowiki><html><span class="part_num">28</span></html>_XP(0830); <nowiki>#</nowiki><html><span class="part_num">27</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">28</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">30</span></html>_XP -> failed!, <nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html>_XP(0831)
*Making medium(SOB, LB amp plate), competent cells (WT & ΔcheZ)
*Making medium(SOB, LB amp plate), competent cells (WT & ΔcheZ)
-
*Miniprep(<nowiki>#</nowiki>3, <nowiki>#</nowiki>20)
+
*Miniprep(<nowiki>#</nowiki><html><span class="part_num">3</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span></html>)
*PCR and Column purification
*PCR and Column purification
-
**<nowiki>#</nowiki>3-14, <nowiki>#</nowiki>3-29, <nowiki>#</nowiki>3-14-5 (Recovery from colony PCR tubes!)
+
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html> (Recovery from colony PCR tubes!)
*Digest
*Digest
-
**<nowiki>#</nowiki>3 -> Failed (Mixed with DMSO!), <nowiki>#</nowiki>20
+
**<nowiki>#</nowiki><html><span class="part_num">3</span></html> -> Failed (Mixed with DMSO!), <nowiki>#</nowiki><html><span class="part_num">20</span></html>
-
**<nowiki>#</nowiki>3-14, <nowiki>#</nowiki>3-29, <nowiki>#</nowiki>3-14-5
+
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>
*Ligation
*Ligation
-
**<nowiki>#</nowiki>3-27, <nowiki>#</nowiki>20-<nowiki>#</nowiki>3-17-5
+
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>
*Transformation
*Transformation
-
**<nowiki>#</nowiki>36 into Nippon Gene comp.
+
**<nowiki>#</nowiki><html><span class="part_num">36</span></html> into Nippon Gene comp.
**Ligation products into Takara comp. (including N.C.)
**Ligation products into Takara comp. (including N.C.)
-
**Titer check(MP product <nowiki>#</nowiki>3 into 0901 WT comp., MP product <nowiki>#</nowiki>20 into 0901 ΔcheZ comp.)
+
**Titer check(MP product <nowiki>#</nowiki><html><span class="part_num">3</span></html> into 0901 WT comp., MP product <nowiki>#</nowiki><html><span class="part_num">20</span></html> into 0901 ΔcheZ comp.)
-
*Mobility assay(<nowiki>#</nowiki>14-5 (km resistance) WT/ΔcheZ cells)
+
*Mobility assay(<nowiki>#</nowiki><html><span class="part_num">14</span>-<span class="part_num">5</span></html> (km resistance) WT/ΔcheZ cells)
<html></div></html>
<html></div></html>
Line 777: Line 780:
*Making medium and reagent(LB 120ml(0.125, 0.25, 0.5% Agar, M9 stock(20×), M9 200ml, Vitamins Solution (100×))
*Making medium and reagent(LB 120ml(0.125, 0.25, 0.5% Agar, M9 stock(20×), M9 200ml, Vitamins Solution (100×))
*Gel extraction -> Digest products were OK but extraction was failed.
*Gel extraction -> Digest products were OK but extraction was failed.
-
**<nowiki>#</nowiki>20_SP, <nowiki>#</nowiki>3-14_XP, <nowiki>#</nowiki>3-14-5_XP, <nowiki>#</nowiki>3-29_XP
+
**<nowiki>#</nowiki><html><span class="part_num">20</span></html>_SP, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span></html>_XP
-
*Colony PCR & Master plate(<nowiki>#</nowiki>20-3-17-5, <nowiki>#</nowiki>3-27)
+
*Colony PCR & Master plate(<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span></html>)
-
*Miniprep(<nowiki>#</nowiki>3, <nowiki>#</nowiki>20, <nowiki>#</nowiki>37, <nowiki>#</nowiki>3-14, <nowiki>#</nowiki>3-29, <nowiki>#</nowiki>3-14-5, <nowiki>#</nowiki>20-3-17-5, <nowiki>#</nowiki>3-27, ΔcheZ)
+
*Miniprep(<nowiki>#</nowiki><html><span class="part_num">3</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span></html>, <nowiki>#</nowiki><html><span class="part_num">37</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span></html>, ΔcheZ)
*Hot-start PCR & Column purification -> PCR products were OK but all purified products were lost.
*Hot-start PCR & Column purification -> PCR products were OK but all purified products were lost.
-
**<nowiki>#</nowiki>20-3-17-5, <nowiki>#</nowiki>3-27 (Recovery from colony PCR tubes)
+
**<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span></html> (Recovery from colony PCR tubes)
-
**<nowiki>#</nowiki>3-14, <nowiki>#</nowiki>3-14-5, <nowiki>#</nowiki>3-29 (from miniprep products)
+
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span></html> (from miniprep products)
*PCR & Column purification
*PCR & Column purification
-
**<nowiki>#</nowiki>20-3-17-5, <nowiki>#</nowiki>3-27, <nowiki>#</nowiki>3-14-5, <nowiki>#</nowiki>3-29 (from miniprep products)
+
**<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span></html> (from miniprep products)
*Digest
*Digest
-
**<nowiki>#</nowiki>3_SP, <nowiki>#</nowiki>20_SP, <nowiki>#</nowiki>37_ES, <nowiki>#</nowiki>3-14_ES, <nowiki>#</nowiki>3-29_ES, <nowiki>#</nowiki>3-14-5_XP, ΔcheZ_E/P (from MP products)
+
**<nowiki>#</nowiki><html><span class="part_num">3</span></html>_SP, <nowiki>#</nowiki><html><span class="part_num">20</span></html>_SP, <nowiki>#</nowiki><html><span class="part_num">37</span></html>_ES, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span></html>_ES, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span></html>_ES, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>_XP, ΔcheZ_E/P (from MP products)
-
**<nowiki>#</nowiki>20-3-17-5_XP, <nowiki>#</nowiki>3-27_ES, <nowiki>#</nowiki>3-14-5_XP, <nowiki>#</nowiki>3-29_ES (from PCR products)
+
**<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span></html>_ES, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span></html>_ES (from PCR products)
<html></div></html>
<html></div></html>
Line 794: Line 797:
*Gel extraction
*Gel extraction
*Ligation & Transformation
*Ligation & Transformation
-
**<nowiki>#</nowiki>20-<nowiki>#</nowiki>3-14-5, <nowiki>#</nowiki>2-<nowiki>#</nowiki>3-14-5, <nowiki>#</nowiki>3-27-<nowiki>#</nowiki>5 into Nippon Gene comp. (+ High Competent Broth 240ul)
+
**<nowiki>#</nowiki><html><span class="part_num">20</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-</html><nowiki>#</nowiki><html><span class="part_num">5</span></html> into Nippon Gene comp. (+ High Competent Broth 240ul)
-
**<nowiki>#</nowiki>3-27-<nowiki>#</nowiki>5, <nowiki>#</nowiki>37-<nowiki>#</nowiki>5 into 0902 WT comp.(+ Takara SOC broth 500ul)
+
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-</html><nowiki>#</nowiki><html><span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">37</span>-</html><nowiki>#</nowiki><html><span class="part_num">5</span></html> into 0902 WT comp.(+ Takara SOC broth 500ul)
*Assays
*Assays
**Mobility assay (.5%, .25%, .125%)
**Mobility assay (.5%, .25%, .125%)
**Asp assay
**Asp assay
-
*Part Cloning (<nowiki>#</nowiki>36)
+
*Part Cloning (<nowiki>#</nowiki><html><span class="part_num">36</span></html>)
<html></div></html>
<html></div></html>
Line 805: Line 808:
==='11/09/04(Sun)===
==='11/09/04(Sun)===
*Hot-start PCR Pre-Mix (4×) preparation (Buffer + dNTPs + THUNDER Taq)
*Hot-start PCR Pre-Mix (4×) preparation (Buffer + dNTPs + THUNDER Taq)
-
*Colony PCR(<nowiki>#</nowiki>20-3-14-5, <nowiki>#</nowiki>3-27-5, <nowiki>#</nowiki>3-29-5, <nowiki>#</nowiki>37-5)
+
*Colony PCR(<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">37</span>-<span class="part_num">5</span></html>)
*Colony Diffusion Assays
*Colony Diffusion Assays
*GFP Live-Imaging Assays
*GFP Live-Imaging Assays
*Ligation & Transformation (retry)  
*Ligation & Transformation (retry)  
-
**<nowiki>#</nowiki>2-<nowiki>#</nowiki>3-14-5, <nowiki>#</nowiki>3-27-<nowiki>#</nowiki>5, <nowiki>#</nowiki>3-29-<nowiki>#</nowiki>5, <nowiki>#</nowiki>37-<nowiki>#</nowiki>5, <nowiki>#</nowiki>20-<nowiki>#</nowiki>28 into 0901 WT comp.
+
**<nowiki>#</nowiki><html><span class="part_num">2</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-</html><nowiki>#</nowiki><html><span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-</html><nowiki>#</nowiki><html><span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">37</span>-</html><nowiki>#</nowiki><html><span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-</html><nowiki>#</nowiki><html><span class="part_num">28</span></html> into 0901 WT comp.
-
**<nowiki>#</nowiki>36 into Takara comp.
+
**<nowiki>#</nowiki><html><span class="part_num">36</span></html> into Takara comp.
-
**<nowiki>#</nowiki>2, <nowiki>#</nowiki>5 as Negative Controls into 0901 WT comp.
+
**<nowiki>#</nowiki><html><span class="part_num">2</span></html>, <nowiki>#</nowiki><html><span class="part_num">5</span></html> as Negative Controls into 0901 WT comp.
-
*Culture for Miniprep(<nowiki>#</nowiki>20-3-14-5, <nowiki>#</nowiki>20)
+
*Culture for Miniprep(<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span></html>)
-
*Digest(<nowiki>#</nowiki>20)
+
*Digest(<nowiki>#</nowiki><html><span class="part_num">20</span></html>)
<html></div></html>
<html></div></html>
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/09/05(Mon)===
==='11/09/05(Mon)===
-
*Colony Diffusion Assays (<nowiki>#</nowiki>2-9) WT,cheZ-
+
*Colony Diffusion Assays (<nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">9</span></html>) WT,cheZ-
-
*Gel extraction(<nowiki>#</nowiki>20)
+
*Gel extraction(<nowiki>#</nowiki><html><span class="part_num">20</span></html>)
*Luciferase Assays
*Luciferase Assays
**Calibrations (Firefly Luciferase standards)
**Calibrations (Firefly Luciferase standards)
**Background Assay
**Background Assay
-
**Dual Luciferase Assay(<nowiki>#</nowiki>20-3-14-5, <nowiki>#</nowiki>20-3-17-5)
+
**Dual Luciferase Assay(<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
-
*Colony PCR(<nowiki>#</nowiki>2-<nowiki>#</nowiki>3-14-5, <nowiki>#</nowiki>3-27-<nowiki>#</nowiki>5, <nowiki>#</nowiki>3-29-<nowiki>#</nowiki>5, <nowiki>#</nowiki>37-<nowiki>#</nowiki>5, <nowiki>#</nowiki>20-<nowiki>#</nowiki>28, <nowiki>#</nowiki>36)
+
*Colony PCR(<nowiki>#</nowiki><html><span class="part_num">2</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-</html><nowiki>#</nowiki><html><span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-</html><nowiki>#</nowiki><html><span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">37</span>-</html><nowiki>#</nowiki><html><span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-</html><nowiki>#</nowiki><html><span class="part_num">28</span></html>, <nowiki>#</nowiki><html><span class="part_num">36</span></html>)
-
*PCR -> clean-up -> Digest(<nowiki>#</nowiki>3-29-<nowiki>#</nowiki>5, <nowiki>#</nowiki>37-5)
+
*PCR -> clean-up -> Digest(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-</html><nowiki>#</nowiki><html><span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">37</span>-<span class="part_num">5</span></html>)
*IPTG-inducted Cell lysis test(IPTG+, LB plate (Positive control))
*IPTG-inducted Cell lysis test(IPTG+, LB plate (Positive control))
-
*Miniprep(<nowiki>#</nowiki>20-3-14-5, <nowiki>#</nowiki>37-5, <nowiki>#</nowiki>36)
+
*Miniprep(<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">37</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">36</span></html>)
*Making M9 -> Asp -> all failed(.125%, .25% Agar)
*Making M9 -> Asp -> all failed(.125%, .25% Agar)
-
*Culture for Miniprep(<nowiki>#</nowiki>2-3-14-5)
+
*Culture for Miniprep(<nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>)
<html></div></html>
<html></div></html>
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/09/06(Tue)===
==='11/09/06(Tue)===
-
*Gel extraction(<nowiki>#</nowiki>37-5, <nowiki>#</nowiki>3-29-5)
+
*Gel extraction(<nowiki>#</nowiki><html><span class="part_num">37</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>)
-
*Miniprep(<nowiki>#</nowiki>2-3-14-5, <nowiki>#</nowiki>20-3-14-5, <nowiki>#</nowiki>20-3-17-5, <nowiki>#</nowiki>36, <nowiki>#</nowiki>3-29-5, <nowiki>#</nowiki>37-5)
+
*Miniprep(<nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">36</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">37</span>-<span class="part_num">5</span></html>)
-
*PCR (retry) (Hot-start & Normal PCR) -> Purification -> Digest(<nowiki>#</nowiki>3-29-5_XP, <nowiki>#</nowiki>37-5_ES)
+
*PCR (retry) (Hot-start & Normal PCR) -> Purification -> Digest(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">37</span>-<span class="part_num">5</span></html>_ES)
*PCR (cloning retry)(lexA, uvrBp)
*PCR (cloning retry)(lexA, uvrBp)
*Ligation & Transformation
*Ligation & Transformation
-
**<nowiki>#</nowiki>3-27-<nowiki>#</nowiki>5, <nowiki>#</nowiki>3-30-<nowiki>#</nowiki>5, <nowiki>#</nowiki>2-3-11-5-pSB1AK3
+
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-</html><nowiki>#</nowiki><html><span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">30</span>-</html><nowiki>#</nowiki><html><span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span>-</html>pSB1AK3
-
**<nowiki>#</nowiki>5, pSB1AK3 (N.C.)
+
**<nowiki>#</nowiki><html><span class="part_num">5</span></html>, pSB1AK3 (N.C.)
-
*Culture for GFP live-imaging (<nowiki>#</nowiki>2-9 WT/cheZ-) -> 4℃ storage
+
*Culture for GFP live-imaging (<nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">9</span></html> WT/cheZ-) -> 4℃ storage
*Ninhydrin sol. preparation (Not dissolvable!) -> 4℃ storage
*Ninhydrin sol. preparation (Not dissolvable!) -> 4℃ storage
<html></div></html>
<html></div></html>
Line 847: Line 850:
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/09/07(Wed)===
==='11/09/07(Wed)===
-
*Colony PCR(<nowiki>#</nowiki>3-27-5, <nowiki>#</nowiki>3-30-5)
+
*Colony PCR(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">30</span>-<span class="part_num">5</span></html>)
-
*Digest(<nowiki>#</nowiki>36_SP)
+
*Digest(<nowiki>#</nowiki><html><span class="part_num">36</span></html>_SP)
-
*Culture from master plate(<nowiki>#</nowiki>36, <nowiki>#</nowiki>3-29-5, <nowiki>#</nowiki>37-5)
+
*Culture from master plate(<nowiki>#</nowiki><html><span class="part_num">36</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">37</span>-<span class="part_num">5</span></html>)
-
*Miniprep(<nowiki>#</nowiki>36, <nowiki>#</nowiki>3-29-5, <nowiki>#</nowiki>37-5, <nowiki>#</nowiki>3-27-5 (failed))
+
*Miniprep(<nowiki>#</nowiki><html><span class="part_num">36</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">37</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html> (failed))
*PCR purification & Digest(lexA_XP, uvrBp_ES)
*PCR purification & Digest(lexA_XP, uvrBp_ES)
-
*Gel extraction(<nowiki>#</nowiki>3-29-5_XP, <nowiki>#</nowiki>37-5_ES, pSB1AK3_EP, <nowiki>#</nowiki>36_SP)
+
*Gel extraction(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">37</span>-<span class="part_num">5</span></html>_ES, pSB1AK3_EP, <nowiki>#</nowiki><html><span class="part_num">36</span></html>_SP)
-
*PCR amplification -> Digest(<nowiki>#</nowiki>3-14-5, <nowiki>#</nowiki>3-29-5, <nowiki>#</nowiki>37-5, <nowiki>#</nowiki>3-27-5)
+
*PCR amplification -> Digest(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">37</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>)
-
*Digest(<nowiki>#</nowiki>36_SP (re), <nowiki>#</nowiki>2_SP, <nowiki>#</nowiki>2-3-17-5_EX, <nowiki>#</nowiki>3-14-5_ES, <nowiki>#</nowiki>34_SP)
+
*Digest(<nowiki>#</nowiki><html><span class="part_num">36</span></html>_SP (re), <nowiki>#</nowiki><html><span class="part_num">2</span></html>_SP, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_EX, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>_ES, <nowiki>#</nowiki><html><span class="part_num">34</span></html>_SP)
-
*Culture for Miniprep(<nowiki>#</nowiki>2, <nowiki>#</nowiki>34, <nowiki>#</nowiki>36, <nowiki>#</nowiki>3-27-5, <nowiki>#</nowiki>3-14-5, <nowiki>#</nowiki>2-3-17-5)
+
*Culture for Miniprep(<nowiki>#</nowiki><html><span class="part_num">2</span></html>, <nowiki>#</nowiki><html><span class="part_num">34</span></html>, <nowiki>#</nowiki><html><span class="part_num">36</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
Ligation & Transformation
Ligation & Transformation
-
**<nowiki>#</nowiki>33-<nowiki>#</nowiki>3-14-5, <nowiki>#</nowiki>34-<nowiki>#</nowiki>3-14-5, <nowiki>#</nowiki>36-<nowiki>#</nowiki>3-14-5, <nowiki>#</nowiki>2-<nowiki>#</nowiki>3-29-5, <nowiki>#</nowiki>20-<nowiki>#</nowiki>3-29-5, <nowiki>#</nowiki>3-<nowiki>#</nowiki>37-5, <nowiki>#</nowiki>2-3-11-5-pSB1AK3, <nowiki>#</nowiki>1-<nowiki>#</nowiki>3-17-5
+
**<nowiki>#</nowiki><html><span class="part_num">33</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">34</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">36</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">37</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span>-</html>pSB1AK3, <nowiki>#</nowiki><html><span class="part_num">1</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>
-
**<nowiki>#</nowiki>33, <nowiki>#</nowiki>34, <nowiki>#</nowiki>36, <nowiki>#</nowiki>1, <nowiki>#</nowiki>2, <nowiki>#</nowiki>20, <nowiki>#</nowiki>3, pSB1AK3 (N.C.)
+
**<nowiki>#</nowiki><html><span class="part_num">33</span></html>, <nowiki>#</nowiki><html><span class="part_num">34</span></html>, <nowiki>#</nowiki><html><span class="part_num">36</span></html>, <nowiki>#</nowiki><html><span class="part_num">1</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span></html>, pSB1AK3 (N.C.)
*Ninhydrin stock solution preparation
*Ninhydrin stock solution preparation
**Asp conc. check (it took 5 min at 80C)
**Asp conc. check (it took 5 min at 80C)
Line 867: Line 870:
==='11/09/08(Thu)===
==='11/09/08(Thu)===
*Making competent cell (WT), LB(amp) plate
*Making competent cell (WT), LB(amp) plate
-
*Frozen stocks(<nowiki>#</nowiki>36, <nowiki>#</nowiki>3-14-5, <nowiki>#</nowiki>3-27-5)
+
*Frozen stocks(<nowiki>#</nowiki><html><span class="part_num">36</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>)
-
*Colony PCR(<nowiki>#</nowiki>33-3-14-5, <nowiki>#</nowiki>34-3-14-5, <nowiki>#</nowiki>36-3-14-5, <nowiki>#</nowiki>20-3-29-5, <nowiki>#</nowiki>2-3-11-5(psB1AK3))
+
*Colony PCR(<nowiki>#</nowiki><html><span class="part_num">33</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">34</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>(psB1AK3))
-
*Miniprep(<nowiki>#</nowiki>2, <nowiki>#</nowiki>34, <nowiki>#</nowiki>36, <nowiki>#</nowiki>2-3-17-5, <nowiki>#</nowiki>3-27-5, <nowiki>#</nowiki>3-14-5)
+
*Miniprep(<nowiki>#</nowiki><html><span class="part_num">2</span></html>, <nowiki>#</nowiki><html><span class="part_num">34</span></html>, <nowiki>#</nowiki><html><span class="part_num">36</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>)
*Gel extraction
*Gel extraction
-
**PCR product: <nowiki>#</nowiki>35_SP, <nowiki>#</nowiki>31_XP, <nowiki>#</nowiki>3-14-5_XP, <nowiki>#</nowiki>3-27-5_XP, <nowiki>#</nowiki>3-29-5_XP, <nowiki>#</nowiki>37-5_XP
+
**PCR product: <nowiki>#</nowiki><html><span class="part_num">35</span></html>_SP, <nowiki>#</nowiki><html><span class="part_num">31</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">37</span>-<span class="part_num">5</span></html>_XP
-
**Miniprep product: <nowiki>#</nowiki>2_SP, <nowiki>#</nowiki>34_SP, <nowiki>#</nowiki>36_SP, <nowiki>#</nowiki>2-3-17-5_EX, <nowiki>#</nowiki>3-14-5_ES
+
**Miniprep product: <nowiki>#</nowiki><html><span class="part_num">2</span></html>_SP, <nowiki>#</nowiki><html><span class="part_num">34</span></html>_SP, <nowiki>#</nowiki><html><span class="part_num">36</span></html>_SP, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_EX, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>_ES
-
*PCR amplification(<nowiki>#</nowiki>34-3-14-5, <nowiki>#</nowiki>36-3-14-5, <nowiki>#</nowiki>20-3-29-5, <nowiki>#</nowiki>20-3-14-5)
+
*PCR amplification(<nowiki>#</nowiki><html><span class="part_num">34</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>)
-
*PCR for Sequencing -> Store at -20C(<nowiki>#</nowiki>33-3-11-5, <nowiki>#</nowiki>34-3-14-5, <nowiki>#</nowiki>20-3-29-5, <nowiki>#</nowiki>37-5)
+
*PCR for Sequencing -> Store at -20C(<nowiki>#</nowiki><html><span class="part_num">33</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">34</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">37</span>-<span class="part_num">5</span></html>)
-
*Ligation(<nowiki>#</nowiki>35_ES-psB1AK3_ES, <nowiki>#</nowiki>31_XP-psB1A2_XP, <nowiki>#</nowiki>31_XP-psB1AK3_XP, <nowiki>#</nowiki>3-<nowiki>#</nowiki>37-5, <nowiki>#</nowiki>36-3-29-5, <nowiki>#</nowiki>3-14-5_ES-<nowiki>#</nowiki>2-3-17-5_EX, <nowiki>#</nowiki>20-<nowiki>#</nowiki>3-27-5)
+
*Ligation(<nowiki>#</nowiki><html><span class="part_num">35</span></html>_ES-psB1AK3_ES, <nowiki>#</nowiki><html><span class="part_num">31</span></html>_XP-psB1A2_XP, <nowiki>#</nowiki><html><span class="part_num">31</span></html>_XP-psB1AK3_XP, <nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">37</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>_ES-<nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_EX, <nowiki>#</nowiki><html><span class="part_num">20</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>)
-
*Culture for Miniprep(<nowiki>#</nowiki>34-3-17-5, <nowiki>#</nowiki>36-3-14-5, <nowiki>#</nowiki>20-3-29-5, <nowiki>#</nowiki>2-3-11-5)
+
*Culture for Miniprep(<nowiki>#</nowiki><html><span class="part_num">34</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>)
*Digest
*Digest
-
**PCR products: <nowiki>#</nowiki>34-3-14-5_ES, <nowiki>#</nowiki>36-3-14-5_ES, <nowiki>#</nowiki>20-3-14-5_ES
+
**PCR products: <nowiki>#</nowiki><html><span class="part_num">34</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>_ES, <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>_ES, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>_ES
**pSB1C3_EP
**pSB1C3_EP
*Asp Gradient test
*Asp Gradient test
Line 886: Line 889:
==='11/09/09(Fri)===
==='11/09/09(Fri)===
*Making competent cell (WT), TB
*Making competent cell (WT), TB
-
*Colony PCR(<nowiki>#</nowiki>35-pSB1AK3 (failed), <nowiki>#</nowiki>31-pSB1AK3, <nowiki>#</nowiki>3-<nowiki>#</nowiki>37-5, <nowiki>#</nowiki>36-<nowiki>#</nowiki>3-29-5, <nowiki>#</nowiki>3-14-5-<nowiki>#</nowiki>2-3-17-5, <nowiki>#</nowiki>20-3-27-5(failed))
+
*Colony PCR(<nowiki>#</nowiki><html><span class="part_num">35</span>-</html>pSB1AK3 (failed), <nowiki>#</nowiki><html><span class="part_num">31</span>-</html>pSB1AK3, <nowiki>#</nowiki><html><span class="part_num">3</span>-</html><nowiki>#</nowiki><html><span class="part_num">37</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">36</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-</html><nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>(failed))
*PCR amplification
*PCR amplification
-
**From Miniprep : <nowiki>#</nowiki>3-14-5, <nowiki>#</nowiki>3-17-5
+
**From Miniprep : <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>
-
**From Colony PCR : <nowiki>#</nowiki>31, <nowiki>#</nowiki>3-37-5, <nowiki>#</nowiki>36-3-29-5, <nowiki>#</nowiki>3-14-5-2-3-17-5 (failed)
+
**From Colony PCR : <nowiki>#</nowiki><html><span class="part_num">31</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html> (failed)
*Digest
*Digest
-
**Miniprep product: <nowiki>#</nowiki>3-11-5_EP
+
**Miniprep product: <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>_EP
-
**PCR-purified product: <nowiki>#</nowiki>3-14-5_EP, <nowiki>#</nowiki>3-17-5_EP, <nowiki>#</nowiki>31_EP, <nowiki>#</nowiki>3-37-5_XP, <nowiki>#</nowiki>36-3-29-5_ES
+
**PCR-purified product: <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>_EP, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_EP, <nowiki>#</nowiki><html><span class="part_num">31</span></html>_EP, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_ES
-
*Miniprep(<nowiki>#</nowiki>34-3-14-5, <nowiki>#</nowiki>36-3-14-5, <nowiki>#</nowiki>20-3-29-5, <nowiki>#</nowiki>2-3-11-5 (pSB1AK3))
+
*Miniprep(<nowiki>#</nowiki><html><span class="part_num">34</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html> (pSB1AK3))
-
*Gel extraction(<nowiki>#</nowiki>34-3-14-5_ES, <nowiki>#</nowiki>36-3-14-5_ES, <nowiki>#</nowiki>20-3-14-5_ES, <nowiki>#</nowiki>3-11-5_EP)
+
*Gel extraction(<nowiki>#</nowiki><html><span class="part_num">34</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>_ES, <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>_ES, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>_ES, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>_EP)
*Ligation
*Ligation
-
**<nowiki>#</nowiki>3-11-5_EP-pSB1C3_EP, <nowiki>#</nowiki>29_XP-pSB1AK3_XP
+
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>_EP-pSB1C3_EP, <nowiki>#</nowiki><html><span class="part_num">29</span></html>_XP-pSB1AK3_XP
-
**<nowiki>#</nowiki>36-3-14-5_ES-<nowiki>#</nowiki>2-3-17-5_EX, <nowiki>#</nowiki>20_SP-<nowiki>#</nowiki>3-27-5_XP(retry)
+
**<nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>_ES-<nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_EX, <nowiki>#</nowiki><html><span class="part_num">20</span></html>_SP-<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>_XP(retry)
*Comp. check
*Comp. check
-
**<nowiki>#</nowiki>2-3-11-5 (pSB1AK3) into 0908 WT comp.
+
**<nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html> (pSB1AK3) into 0908 WT comp.
-
**<nowiki>#</nowiki>20-3-29-5 into 0909 WT comp.
+
**<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html> into 0909 WT comp.
-
**<nowiki>#</nowiki>20-3-29-5 into 0802 cheZ- comp.
+
**<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html> into 0802 cheZ- comp.
*Part cloning(BBa_J04450 (IPTG-induced RFP reporter on pSB1C3))
*Part cloning(BBa_J04450 (IPTG-induced RFP reporter on pSB1C3))
-
*Culture for Miniprep(<nowiki>#</nowiki>31-pSB1AK3, <nowiki>#</nowiki>3-37-5, <nowiki>#</nowiki>36-3-29-5, <nowiki>#</nowiki>3-14-5-2-3-17-5)
+
*Culture for Miniprep(<nowiki>#</nowiki><html><span class="part_num">31</span>-</html>pSB1AK3, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
-
*UV irradiation -> Luciferase Assay(<nowiki>#</nowiki>24-3-17-5, <nowiki>#</nowiki>34-3-17-5)
+
*UV irradiation -> Luciferase Assay(<nowiki>#</nowiki><html><span class="part_num">24</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">34</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
*Asp Gradient test
*Asp Gradient test
-
*CleanSEQ -> Sequencing(<nowiki>#</nowiki>33-3-11-5, <nowiki>#</nowiki>34-3-14-5, <nowiki>#</nowiki>20-3-29-5, <nowiki>#</nowiki>37-5)
+
*CleanSEQ -> Sequencing(<nowiki>#</nowiki><html><span class="part_num">33</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">34</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">37</span>-<span class="part_num">5</span></html>)
<html></div></html>
<html></div></html>
Line 912: Line 915:
==='11/09/10(Sat)===
==='11/09/10(Sat)===
*Culture plate storage (4℃)
*Culture plate storage (4℃)
-
*Miniprep(<nowiki>#</nowiki>31-pSB1AK3, <nowiki>#</nowiki>3-37-5, <nowiki>#</nowiki>36-3-29-5, <nowiki>#</nowiki>3-14-5-2-3-17-5)
+
*Miniprep(<nowiki>#</nowiki><html><span class="part_num">31</span>-</html>pSB1AK3, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
-
*Gel extraction(<nowiki>#</nowiki>3-14-5_EP, <nowiki>#</nowiki>3-17-5_EP, <nowiki>#</nowiki>31_EP, <nowiki>#</nowiki>3-37-5_XP, <nowiki>#</nowiki>36-3-29-5_ES)
+
*Gel extraction(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>_EP, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_EP, <nowiki>#</nowiki><html><span class="part_num">31</span></html>_EP, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_ES)
-
*Colony PCR(<nowiki>#</nowiki>36-3-14-5-<nowiki>#</nowiki>2-3-17-5 (Maybe Ligation was FAILURE , estimated length is 1000-1500 by electrophoresis))
+
*Colony PCR(<nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-</html><nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html> (Maybe Ligation was FAILURE , estimated length is 1000-1500 by electrophoresis))
-
*Ligation(<nowiki>#</nowiki>20_SP-<nowiki>#</nowiki>3-37-5_XP)
+
*Ligation(<nowiki>#</nowiki><html><span class="part_num">20</span></html>_SP-<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>_XP)
<html></div></html>
<html></div></html>
Line 921: Line 924:
==='11/09/11(Sun)===
==='11/09/11(Sun)===
*Digest from Miniprep products
*Digest from Miniprep products
-
**<nowiki>#</nowiki>3-14-5-2-3-17-5_XP
+
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_XP
-
**<nowiki>#</nowiki>36-3-14-5_SP, <nowiki>#</nowiki>20_SP -> O.N.
+
**<nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>_SP, <nowiki>#</nowiki><html><span class="part_num">20</span></html>_SP -> O.N.
*Colony PCR
*Colony PCR
-
**<nowiki>#</nowiki>3-11-5 (pSB1C3), <nowiki>#</nowiki>29 (pSB1AK3), BBa_J04450 (pSB1C3)
+
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html> (pSB1C3), <nowiki>#</nowiki><html><span class="part_num">29</span></html> (pSB1AK3), BBa_J04450 (pSB1C3)
-
**<nowiki>#</nowiki>20-<nowiki>#</nowiki>3-27-5(failed), <nowiki>#</nowiki>20-<nowiki>#</nowiki>3-37-5, <nowiki>#</nowiki>36-<nowiki>#</nowiki>3-14-5-2-3-17-5
+
**<nowiki>#</nowiki><html><span class="part_num">20</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>(failed), <nowiki>#</nowiki><html><span class="part_num">20</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">36</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>
-
*Gel extraction(<nowiki>#</nowiki>3-14-5-2-3-17-5_XP)
+
*Gel extraction(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_XP)
*Culture for Miniprep
*Culture for Miniprep
-
**From Master plate : <nowiki>#</nowiki>20-3-37-5, <nowiki>#</nowiki>29 (pSB1AK3), BBa_J04450 (pSB1C3)
+
**From Master plate : <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">29</span></html> (pSB1AK3), BBa_J04450 (pSB1C3)
-
**From plate stock : <nowiki>#</nowiki>20
+
**From plate stock : <nowiki>#</nowiki><html><span class="part_num">20</span></html>
-
**From frozen stock : <nowiki>#</nowiki>2-3-17-5
+
**From frozen stock : <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>
-
*Miniprep( <nowiki>#</nowiki>36-3-14-5-2-3-17-5)
+
*Miniprep( <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
-
*Ligation(<nowiki>#</nowiki>20_SP-<nowiki>#</nowiki>3-14-5-2-3-17-5_XP, <nowiki>#</nowiki>1_SP-<nowiki>#</nowiki>3-14-5-2-3-17-5_XP)
+
*Ligation(<nowiki>#</nowiki><html><span class="part_num">20</span></html>_SP-<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">1</span></html>_SP-<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_XP)
*Direct Ligation
*Direct Ligation
-
**i.PCR amplification -> Digest for >1h -> Heat kill for 20min (<nowiki>#</nowiki>20-3-37-5_XP, <nowiki>#</nowiki>36-3-14-5-2-3-17-5_ES)
+
**i.PCR amplification -> Digest for >1h -> Heat kill for 20min (<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_ES)
-
**ii.Digest for >1h -> Heat kill for 20min (<nowiki>#</nowiki>36-3-14-5-2-3-17-5_SP, pSB1A3_EP, <nowiki>#</nowiki>2-3-17-5_EX)
+
**ii.Digest for >1h -> Heat kill for 20min (<nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_SP, pSB1A3_EP, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_EX)
-
**iii.Ligation for 30min (<nowiki>#</nowiki>20-3-14-5_ES-<nowiki>#</nowiki>2-3-17-5_EX, <nowiki>#</nowiki>36-3-14-5-2-3-17-5_SP-<nowiki>#</nowiki>20-3-37-5_XP, <nowiki>#</nowiki>36-3-14-5-2-3-17-5_ES-<nowiki>#</nowiki>20-3-37-5_XP-pSB1A3_EP)
+
**iii.Ligation for 30min (<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>_ES-<nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_EX, <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_SP-<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_ES-<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>_XP-pSB1A3_EP)
**iv.Transformation
**iv.Transformation
*Making 0.125% agar LB amp plate (IPTG 1, 10, 30, 100uM)
*Making 0.125% agar LB amp plate (IPTG 1, 10, 30, 100uM)
*Cell Diffusion Assay
*Cell Diffusion Assay
-
**<nowiki>#</nowiki>20-3-29-5 WT (IPTG 1, 10, 30, 100uM)
+
**<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html> WT (IPTG 1, 10, 30, 100uM)
-
**<nowiki>#</nowiki>20-3-29-5 cheZ-/- (IPTG 1, 10, 30, 100uM)
+
**<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html> cheZ-/- (IPTG 1, 10, 30, 100uM)
**WT
**WT
**cheZ-/-
**cheZ-/-
Line 949: Line 952:
==='11/9/12 (Mon)===
==='11/9/12 (Mon)===
*Colony PCR
*Colony PCR
-
**<nowiki>#</nowiki>20-<nowiki>#</nowiki>3-14-5-2-3-17-5
+
**<nowiki>#</nowiki><html><span class="part_num">20</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>
-
**<nowiki>#</nowiki>20-3-14-5-<nowiki>#</nowiki>2-3-17-5
+
**<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-</html><nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>
-
**<nowiki>#</nowiki>1-<nowiki>#</nowiki>3-14-5-2-3-17-5
+
**<nowiki>#</nowiki><html><span class="part_num">1</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>
-
**<nowiki>#</nowiki>36-3-14-5-2-3-17-5-<nowiki>#</nowiki>20-3-37-5 (no colony)
+
**<nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span>-</html><nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html> (no colony)
-
**<nowiki>#</nowiki>36-3-14-5-2-3-17-5-<nowiki>#</nowiki>20-3-37-5-pSB1A3 (PCR failed?)
+
**<nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span>-</html><nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span>-</html>pSB1A3 (PCR failed?)
-
Note: <nowiki>#</nowiki>20-3-37-5 colonies on the master-plate are RED. Maybe <nowiki>#</nowiki>2-3-11-5...<br>
+
Note: <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html> colonies on the master-plate are RED. Maybe <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>...<br>
-
Note: <nowiki>#</nowiki>36-3-14-5-2-3-17-5-<nowiki>#</nowiki>20-3-37-5-pSB1A3 PCR amplification doesn't work. Seems failed extension...
+
Note: <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span>-</html><nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span>-</html>pSB1A3 PCR amplification doesn't work. Seems failed extension...
*PCR amplification
*PCR amplification
-
**From master plate : <nowiki>#</nowiki>20-3-37-5
+
**From master plate : <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>
-
**From 09/11 Miniprep product : <nowiki>#</nowiki>36-3-14-5-2-3-17-5
+
**From 09/11 Miniprep product : <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>
-
Note: Only one <nowiki>#</nowiki>20-3-37-5 colony is NOT RED.
+
Note: Only one <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html> colony is NOT RED.
-
*Miniprep(<nowiki>#</nowiki>20, <nowiki>#</nowiki>29 (pSB1AK3), <nowiki>#</nowiki>2-3-17-5, <nowiki>#</nowiki>2-3-37-5, BBa_J04450 (pSB1C3))
+
*Miniprep(<nowiki>#</nowiki><html><span class="part_num">20</span></html>, <nowiki>#</nowiki><html><span class="part_num">29</span></html> (pSB1AK3), <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>, BBa_J04450 (pSB1C3))
*Digest
*Digest
-
**From PCR : <nowiki>#</nowiki>36-3-14-5-2-3-17-5_ES, <nowiki>#</nowiki>20-3-37-5_XP
+
**From PCR : <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_ES, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>_XP
**pSB1A3_EP, pSB1C3_EP
**pSB1A3_EP, pSB1C3_EP
-
**From Miniprep : <nowiki>#</nowiki>20-3-37-5_XP
+
**From Miniprep : <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>_XP
-
**J04450_EP, *<nowiki>#</nowiki>2-9_XP, <nowiki>#</nowiki>20-3-29-5_ES
+
**J04450_EP, *<nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">9</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_ES
*Gel extraction
*Gel extraction
-
**<nowiki>#</nowiki>20_SP, <nowiki>#</nowiki>36-3-14-5_SP
+
**<nowiki>#</nowiki><html><span class="part_num">20</span></html>_SP, <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>_SP
-
**From OverNight digests : <nowiki>#</nowiki>2-3-17-5_EX
+
**From OverNight digests : <nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_EX
-
**<nowiki>#</nowiki>36-3-14-5-2-3-17-5_ES
+
**<nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_ES
-
**From today's PCR digests : <nowiki>#</nowiki>20-3-37-5_XP
+
**From today's PCR digests : <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>_XP
-
*Culture for Miniprep(<nowiki>#</nowiki>20-3-37-5 from 0911 master plate)
+
*Culture for Miniprep(<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html> from 0911 master plate)
-
*Dual Luciferase Assay(<nowiki>#</nowiki>20-3-14-5-2-3-17-5, <nowiki>#</nowiki>1-3-14-5-2-3-17-5, <nowiki>#</nowiki>36-3-14-5-2-3-17-5-20-3-37-5-pSB1A3)
+
*Dual Luciferase Assay(<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">1</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span>-<span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span>-</html>pSB1A3)
*Ligation
*Ligation
-
**<nowiki>#</nowiki>36-3-14-5-2-3-17-5_ES-<nowiki>#</nowiki>20-3-37-5_XP-pSB1A3_EP
+
**<nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_ES-<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>_XP-pSB1A3_EP
-
**<nowiki>#</nowiki>36-3-14-5-2-3-17-5_ES-<nowiki>#</nowiki>20-3-37-5_XP-pSB1C3_EP
+
**<nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_ES-<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>_XP-pSB1C3_EP
-
**<nowiki>#</nowiki>3-27-5_XP-<nowiki>#</nowiki>20_SP
+
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>_XP-<nowiki>#</nowiki><html><span class="part_num">20</span></html>_SP
-
**<nowiki>#</nowiki>36-3-29-5_ES-<nowiki>#</nowiki>20-3-37-5_XP-pSB1AK3_EP
+
**<nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_ES-<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>_XP-pSB1AK3_EP
<html></div></html>
<html></div></html>
Line 983: Line 986:
==='11/9/13 (Tue)===
==='11/9/13 (Tue)===
*Colony PCR
*Colony PCR
-
**<nowiki>#</nowiki>36-3-14-5-2-3-17-5_ES-<nowiki>#</nowiki>20-3-37-5_XP-pSB1A3_EP
+
**<nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_ES-<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>_XP-pSB1A3_EP
-
**<nowiki>#</nowiki>36-3-14-5-2-3-17-5_ES-<nowiki>#</nowiki>20-3-37-5_XP-pSB1C3_EP (failed)
+
**<nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_ES-<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>_XP-pSB1C3_EP (failed)
-
**<nowiki>#</nowiki>3-27-5_XP-<nowiki>#</nowiki>20_SP (failed)
+
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>_XP-<nowiki>#</nowiki><html><span class="part_num">20</span></html>_SP (failed)
-
**<nowiki>#</nowiki>36-3-29-5_ES-<nowiki>#</nowiki>20-3-37-5_XP-pSB1AK3_EP
+
**<nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_ES-<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>_XP-pSB1AK3_EP
*Culture for Luciferase Assay
*Culture for Luciferase Assay
-
**<nowiki>#</nowiki>36-3-14-5-2-3-17-5-20-3-37-5(pSB1A3, pSB1C3) (IPTG 0, 1, 10, 38, 100uM)
+
**<nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span>-<span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>(pSB1A3, pSB1C3) (IPTG 0, 1, 10, 38, 100uM)
-
**<nowiki>#</nowiki>20-3-14-5-2-3-17-5 (IPTG 0, 1, 10, 38, 100uM)
+
**<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html> (IPTG 0, 1, 10, 38, 100uM)
-
**<nowiki>#</nowiki>1-3-14-5-2-3-17-5 (IPTG 0uM)<br>
+
**<nowiki>#</nowiki><html><span class="part_num">1</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html> (IPTG 0uM)<br>
Note: Seemes OD600 should NOT exceed 0.4~0.5, or intracellular luciferase will be saturated.
Note: Seemes OD600 should NOT exceed 0.4~0.5, or intracellular luciferase will be saturated.
*Miniprep.
*Miniprep.
-
**<nowiki>#</nowiki>20-3-14-5-2-3-17-5
+
**<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>
-
**<nowiki>#</nowiki>1-3-14-5-2-3-17-5
+
**<nowiki>#</nowiki><html><span class="part_num">1</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>
-
**<nowiki>#</nowiki>20-3-37-5 (from a non-red colony on the master plate)
+
**<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html> (from a non-red colony on the master plate)
*Gel extraction
*Gel extraction
-
**<nowiki>#</nowiki>20-3-37-5_XP (from non-red colony)
+
**<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>_XP (from non-red colony)
-
**<nowiki>#</nowiki>20-3-29-5_ES(failed)
+
**<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_ES(failed)
-
**<nowiki>#</nowiki>2-9_XP, pSB1C3_EP
+
**<nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">9</span></html>_XP, pSB1C3_EP
-
Note: <nowiki>#</nowiki>20-3-29-5 was not cut!
+
Note: <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html> was not cut!
*Dual Luciferase Assay
*Dual Luciferase Assay
-
**<nowiki>#</nowiki>1-3-14-5-2-3-17-5 (IPTG 0uM)
+
**<nowiki>#</nowiki><html><span class="part_num">1</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html> (IPTG 0uM)
-
**<nowiki>#</nowiki>20-3-14-5-2-3-17-5 (IPTG 0, 1, 10, 38, 100uM)
+
**<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html> (IPTG 0, 1, 10, 38, 100uM)
-
**<nowiki>#</nowiki>36-3-14-5-2-3-17-5-20-3-37-5 (<nowiki>#</nowiki>2-3-11-5?) (IPTG 0, 1, 10, 38, 100uM)
+
**<nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span>-<span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html> (<nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>?) (IPTG 0, 1, 10, 38, 100uM)
*Sequencing preparation
*Sequencing preparation
-
**<nowiki>#</nowiki>20-3-37-5 (from a non-red colony on the master plate)
+
**<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html> (from a non-red colony on the master plate)
-
**<nowiki>#</nowiki>20-3-29-5
+
**<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>
-
**<nowiki>#</nowiki>3-27-5
+
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>
-
**<nowiki>#</nowiki>31
+
**<nowiki>#</nowiki><html><span class="part_num">31</span></html>
*Digest
*Digest
-
**<nowiki>#</nowiki>20_E
+
**<nowiki>#</nowiki><html><span class="part_num">20</span></html>_E
-
**<nowiki>#</nowiki>3-29-5_S (from MP products) (cut check)
+
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_S (from MP products) (cut check)
*Ligation
*Ligation
-
**<nowiki>#</nowiki>20_SP-<nowiki>#</nowiki>3-27-5_XP
+
**<nowiki>#</nowiki><html><span class="part_num">20</span></html>_SP-<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>_XP
*Transformation
*Transformation
-
**<nowiki>#</nowiki>3-14-5-2-3-17-5
+
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>
-
**<nowiki>#</nowiki>1-3-14-5-2-3-17-5
+
**<nowiki>#</nowiki><html><span class="part_num">1</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>
-
**<nowiki>#</nowiki>20-3-14-5-2-3-17-5 (for plate stock)
+
**<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html> (for plate stock)
-
**<nowiki>#</nowiki>20-3-37-5 (from a non-red colony on the master plate) (for plasmid check)
+
**<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html> (from a non-red colony on the master plate) (for plasmid check)
<html></div></html>
<html></div></html>
Line 1,025: Line 1,028:
==='11/09/14(Wed)===
==='11/09/14(Wed)===
*Colony PCR
*Colony PCR
-
**<nowiki>#</nowiki>3-27-5_XP-<nowiki>#</nowiki>20_SP (failed)
+
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>_XP-<nowiki>#</nowiki><html><span class="part_num">20</span></html>_SP (failed)
*Culture for Luciferase Assay (from plate stocks)
*Culture for Luciferase Assay (from plate stocks)
-
**<nowiki>#</nowiki>20-3-14-5-2-3-17-5 (IPTG 0, 1, 10, 38, 100uM)
+
**<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html> (IPTG 0, 1, 10, 38, 100uM)
-
**<nowiki>#</nowiki>1-3-14-5-2-3-17-5 (IPTG 0uM)
+
**<nowiki>#</nowiki><html><span class="part_num">1</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html> (IPTG 0uM)
*Digest
*Digest
-
**<nowiki>#</nowiki>3-14-5, <nowiki>#</nowiki>3-17-5, <nowiki>#</nowiki>3-14-5-2-3-17-5, <nowiki>#</nowiki>1-3-14-5-2-3-17-5, <nowiki>#</nowiki>20-3-14-5-2-3-17-5 (EPcut)
+
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">1</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html> (EPcut)
-
**<nowiki>#</nowiki>20_SP
+
**<nowiki>#</nowiki><html><span class="part_num">20</span></html>_SP
*Culture for Miniprep.
*Culture for Miniprep.
-
**From plate stock : <nowiki>#</nowiki>20, <nowiki>#</nowiki>20-3-14-5-2-3-17-5, <nowiki>#</nowiki>3-14-5-2-3-17-5, <nowiki>#</nowiki>3-17-5  
+
**From plate stock : <nowiki>#</nowiki><html><span class="part_num">20</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>
-
**From 09/11 master plate : <nowiki>#</nowiki>3-11-5 (pSB1C3)
+
**From 09/11 master plate : <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html> (pSB1C3)
-
*Part cloning(<nowiki>#</nowiki>19: BBa_R0010)
+
*Part cloning(<nowiki>#</nowiki><html><span class="part_num">19</span></html>: BBa_R0010)
*Ligation
*Ligation
-
**<nowiki>#</nowiki>20_SP-<nowiki>#</nowiki>3-29-5_XP
+
**<nowiki>#</nowiki><html><span class="part_num">20</span></html>_SP-<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_XP
-
**<nowiki>#</nowiki>36-3-14-5-2-3-17-5_ES-<nowiki>#</nowiki>20-3-37-5_XP-pSB1C3
+
**<nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_ES-<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>_XP-pSB1C3
*Making WT competent cell
*Making WT competent cell
*Dual Luciferase Assay
*Dual Luciferase Assay
-
**<nowiki>#</nowiki>1-3-14-5-2-3-17-5 (IPTG 0uM)
+
**<nowiki>#</nowiki><html><span class="part_num">1</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html> (IPTG 0uM)
-
**<nowiki>#</nowiki>20-3-14-5-2-3-17-5 (IPTG 0, 1, 10, 38, 100uM)
+
**<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html> (IPTG 0, 1, 10, 38, 100uM)
*Asp TLC Assay
*Asp TLC Assay
*Asp Diffusion Assay (0.25% Agar)
*Asp Diffusion Assay (0.25% Agar)
*Fumaric Acid (Fum) Sol. preparation
*Fumaric Acid (Fum) Sol. preparation
*Sequencing
*Sequencing
-
**①F⑤R<nowiki>#</nowiki>20-3-37-5 (from a non-red colony on the master plate)
+
**①F⑤R<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html> (from a non-red colony on the master plate)
-
**②F⑥R<nowiki>#</nowiki>20-3-29-5 (<nowiki>#</nowiki>2-3-11-5?)
+
**②F⑥R<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html> (<nowiki>#</nowiki><html><span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>?)
-
**③F⑦R<nowiki>#</nowiki>3-27-5
+
**③F⑦R<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>
-
**④F⑧R<nowiki>#</nowiki>31
+
**④F⑧R<nowiki>#</nowiki><html><span class="part_num">31</span></html>
<html></div></html>
<html></div></html>
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/09/15(Thu)===
==='11/09/15(Thu)===
-
*Colony PCR(<nowiki>#</nowiki>19, <nowiki>#</nowiki>36-3-14-5-2-3-17-5-20-3-37-5 (failed), <nowiki>#</nowiki>20-3-29-5)
+
*Colony PCR(<nowiki>#</nowiki><html><span class="part_num">19</span></html>, <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span>-<span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html> (failed), <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>)
-
*Miniprep(<nowiki>#</nowiki>20, <nowiki>#</nowiki>20-3-14-5-2-3-17-5, <nowiki>#</nowiki>3-14-5-2-3-17-5, <nowiki>#</nowiki>3-17-5)
+
*Miniprep(<nowiki>#</nowiki><html><span class="part_num">20</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
*Gel extraction
*Gel extraction
-
**<nowiki>#</nowiki>3-14-5(failed), <nowiki>#</nowiki>3-17-5, <nowiki>#</nowiki>3-14-5-2-3-17-5, <nowiki>#</nowiki>1-3-14-5-2-3-17-5, <nowiki>#</nowiki>20-3-14-5-2-3-17-5 (EPcut); <nowiki>#</nowiki>20_SP
+
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>(failed), <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">1</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html> (EPcut); <nowiki>#</nowiki><html><span class="part_num">20</span></html>_SP
*Making WT competent cell
*Making WT competent cell
*Making LB plates(amp:39, cm:18)
*Making LB plates(amp:39, cm:18)
-
*Culture for Miniprep(<nowiki>#</nowiki>3-11-5 (pSB1C3), <nowiki>#</nowiki>20-3-29-5, <nowiki>#</nowiki>19)
+
*Culture for Miniprep(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html> (pSB1C3), <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">19</span></html>)
*Ligation
*Ligation
-
**sulAp(ES)-<nowiki>#</nowiki>3-14-5-2-3-17-5_XP-pSB1C3_PCR digest(EP)
+
**sulAp(ES)-<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_XP-pSB1C3_PCR digest(EP)
-
**recNp(ES)-<nowiki>#</nowiki>3-14-5-2-3-17-5_XP-pSB1C3_PCR digest(EP)
+
**recNp(ES)-<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_XP-pSB1C3_PCR digest(EP)
-
**<nowiki>#</nowiki>36-3-14-5-2-3-17-5_ES-<nowiki>#</nowiki>20-3-37-5_XP-pSB1C3_PCR digest(EP)
+
**<nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_ES-<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>_XP-pSB1C3_PCR digest(EP)
-
**<nowiki>#</nowiki>3-27-5_XP-<nowiki>#</nowiki>20_SP
+
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>_XP-<nowiki>#</nowiki><html><span class="part_num">20</span></html>_SP
-
**<nowiki>#</nowiki>3-17-5_EP-pSB1C3_PCR digest(EP)
+
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_EP-pSB1C3_PCR digest(EP)
-
**<nowiki>#</nowiki>3-14-5_EP-pSB1C3_PCR digest(EP)
+
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>_EP-pSB1C3_PCR digest(EP)
-
**<nowiki>#</nowiki>3-14-5-2-3-17-5_EP-pSB1C3_PCR digest(EP)
+
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_EP-pSB1C3_PCR digest(EP)
-
**<nowiki>#</nowiki>20-3-14-5-2-3-17-5_EP-pSB1C3_PCR digest(EP)
+
**<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_EP-pSB1C3_PCR digest(EP)
-
*Culture for Dual Luciferase Assay(<nowiki>#</nowiki>20-3-14-5-2-3-17-5)
+
*Culture for Dual Luciferase Assay(<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
*TLC check (LB, M9)
*TLC check (LB, M9)
*Cell Diffusion Assay
*Cell Diffusion Assay
Line 1,082: Line 1,085:
==='11/09/16(Fri)===
==='11/09/16(Fri)===
*Colony PCR
*Colony PCR
-
**sulAp(ES)-<nowiki>#</nowiki>3-14-5-2-3-17-5_XP-pSB1C3_PCR digest(EP),
+
**sulAp(ES)-<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_XP-pSB1C3_PCR digest(EP),
-
**recNp(ES)-<nowiki>#</nowiki>3-14-5-2-3-17-5_XP-pSB1C3_PCR digest(EP),
+
**recNp(ES)-<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_XP-pSB1C3_PCR digest(EP),
-
**<nowiki>#</nowiki>36-3-14-5-2-3-17-5_ES-<nowiki>#</nowiki>20-3-37-5_XP-pSB1C3_PCR digest(EP),
+
**<nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_ES-<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>_XP-pSB1C3_PCR digest(EP),
-
**<nowiki>#</nowiki>3-27-5_XP-<nowiki>#</nowiki>20_SP,
+
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>_XP-<nowiki>#</nowiki><html><span class="part_num">20</span></html>_SP,
-
**<nowiki>#</nowiki>3-17-5_EP-pSB1C3_PCR digest(EP),
+
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_EP-pSB1C3_PCR digest(EP),
-
**<nowiki>#</nowiki>3-14-5_EP-pSB1C3_PCR digest(EP),
+
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>_EP-pSB1C3_PCR digest(EP),
-
**<nowiki>#</nowiki>3-14-5-2-3-17-5_EP-pSB1C3_PCR digest(EP),
+
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_EP-pSB1C3_PCR digest(EP),
-
**<nowiki>#</nowiki>20-3-14-5-2-3-17-5_EP-pSB1C3_PCR digest(EP)
+
**<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_EP-pSB1C3_PCR digest(EP)
-
**<nowiki>#</nowiki>36-3-14-5-2-3-17-5_ES-<nowiki>#</nowiki>20-3-37-5_XP-pSB1C3_PCR digest(EP) (retry; non-red colonies)
+
**<nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_ES-<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>_XP-pSB1C3_PCR digest(EP) (retry; non-red colonies)
-
*Miniprep(<nowiki>#</nowiki>3-11-5 (pSB1C3), <nowiki>#</nowiki>20-3-29-5, <nowiki>#</nowiki>19)
+
*Miniprep(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html> (pSB1C3), <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">19</span></html>)
-
*PCR amplification(<nowiki>#</nowiki>3-27-5, <nowiki>#</nowiki>36-3-14-5-2-3-17-5, pSB1C3)
+
*PCR amplification(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, pSB1C3)
-
*Sequencing prep(<nowiki>#</nowiki>20-3-29-5)
+
*Sequencing prep(<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>)
*Making agarose gel, WT competent cell,  
*Making agarose gel, WT competent cell,  
*Electrophoresis check(pSB1C3_PCR)
*Electrophoresis check(pSB1C3_PCR)
-
*Digest(pSB1C3_ES, pSB1C3_EP, <nowiki>#</nowiki>3-27-5_XP, <nowiki>#</nowiki>36-3-14-5-2-3-17-5_ES)
+
*Digest(pSB1C3_ES, pSB1C3_EP, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_ES)
*Dual luciferase assay
*Dual luciferase assay
-
**<nowiki>#</nowiki>20-3-14-5-2-3-17-5 (1h, 1.5h culture)(IPTG 0, 1, 10, 100uM)(2.5mL)
+
**<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html> (1h, 1.5h culture)(IPTG 0, 1, 10, 100uM)(2.5mL)
*Fumaric Acid (Fum) Sol. preparation
*Fumaric Acid (Fum) Sol. preparation
**TLC test
**TLC test
Line 1,108: Line 1,111:
==='11/09/17(Sat)===
==='11/09/17(Sat)===
*Colony PCR (retry)
*Colony PCR (retry)
-
**<nowiki>#</nowiki>3-14-5, <nowiki>#</nowiki>3-17-5 (pSB1C3)
+
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html> (pSB1C3)
-
**<nowiki>#</nowiki>36-3-14-5-2-3-17-5-20-3-37-5
+
**<nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span>-<span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>
-
*Culture for Miniprep(<nowiki>#</nowiki>3-14-5, <nowiki>#</nowiki>3-17-5 (pSB1C3))
+
*Culture for Miniprep(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html> (pSB1C3))
-
*Ligation(sulAp-pSB1C3, recNp-pSB1C3, <nowiki>#</nowiki>20-<nowiki>#</nowiki>3-27-5)
+
*Ligation(sulAp-pSB1C3, recNp-pSB1C3, <nowiki>#</nowiki><html><span class="part_num">20</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>)
-
*Transformation(<nowiki>#</nowiki>20-3-29-5 into cheZ-)
+
*Transformation(<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html> into cheZ-)
*Making medium(.25% Agar. M9& LB each 20 plates)
*Making medium(.25% Agar. M9& LB each 20 plates)
*Cell diffusion assay
*Cell diffusion assay
Line 1,119: Line 1,122:
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/09/18(Sun)===
==='11/09/18(Sun)===
-
*Colony PCR & Culture for Miniprep(<nowiki>#</nowiki>36-3-14-5-2-3-17-5-20-3-37-5, <nowiki>#</nowiki>33-3-14-5-2-3-17-5)
+
*Colony PCR & Culture for Miniprep(<nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span>-<span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">33</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
-
*Miniprep(<nowiki>#</nowiki>3-14-5, <nowiki>#</nowiki>3-17-5 (pSB1C3))
+
*Miniprep(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html> (pSB1C3))
-
*Ligation(<nowiki>#</nowiki>20-<nowiki>#</nowiki>3-27-5, recNp-<nowiki>#</nowiki>3-14-5-2-3-17-5, sulAp-pSB1C3, recNp-pSB1C3, <nowiki>#</nowiki>31-pSB1C3)
+
*Ligation(<nowiki>#</nowiki><html><span class="part_num">20</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>, recNp-<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, sulAp-pSB1C3, recNp-pSB1C3, <nowiki>#</nowiki><html><span class="part_num">31</span>-</html>pSB1C3)
*Cell diffusion assay
*Cell diffusion assay
**Innoculate colony(WT / cheZ-/- / cheZ-/- + cheZ plasmid)
**Innoculate colony(WT / cheZ-/- / cheZ-/- + cheZ plasmid)
Line 1,129: Line 1,132:
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/09/19(Mon)===
==='11/09/19(Mon)===
-
*Culture for Dual luciferase assay(<nowiki>#</nowiki>36-3-14-5-2-3-17-5-20-3-37-5, <nowiki>#</nowiki>33-3-14-5-2-3-17-5, <nowiki>#</nowiki>1-3-14-5-2-3-17-5, <nowiki>#</nowiki>20-3-14-5-2-3-17-5)
+
*Culture for Dual luciferase assay(<nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span>-<span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">33</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">1</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
-
*Miniprep(<nowiki>#</nowiki>36-3-14-5-2-3-17-5-20-3-37-5, <nowiki>#</nowiki>33-3-14-5-2-3-17-5)
+
*Miniprep(<nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span>-<span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">33</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
-
*Ligation(<nowiki>#</nowiki>24-<nowiki>#</nowiki>3-14-5-2-3-17-5, <nowiki>#</nowiki>20-<nowiki>#</nowiki>3-27-5)
+
*Ligation(<nowiki>#</nowiki><html><span class="part_num">24</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>)
-
*Sequencing(<nowiki>#</nowiki>36-3-14-5-2-3-17-5-20-3-37-5, <nowiki>#</nowiki>33-3-14-5-2-3-17-5, <nowiki>#</nowiki>20-3-29-5)
+
*Sequencing(<nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span>-<span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">33</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>)
-
*Dual luciferase assay(<nowiki>#</nowiki>1~, <nowiki>#</nowiki>20~, <nowiki>#</nowiki>36~, <nowiki>#</nowiki>33~ )
+
*Dual luciferase assay(<nowiki>#</nowiki><html><span class="part_num">1</span></html>~, <nowiki>#</nowiki><html><span class="part_num">20</span></html>~, <nowiki>#</nowiki><html><span class="part_num">36</span></html>~, <nowiki>#</nowiki><html><span class="part_num">33</span></html>~ )
*Asp racing assay(Innoculate colonies)
*Asp racing assay(Innoculate colonies)
*GFP imaging assay<html></div></html>
*GFP imaging assay<html></div></html>
Line 1,139: Line 1,142:
<html><div class="labDay"></html>
<html><div class="labDay"></html>
==='11/09/20(Tue)===
==='11/09/20(Tue)===
-
*colony PCR(20-3-27-5, <nowiki>#</nowiki>24-3-14-5-2-3-17-5)
+
*colony PCR(20-3-27-5, <nowiki>#</nowiki><html><span class="part_num">24</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
-
*Gel extraction(<nowiki>#</nowiki>3-27-5_XP)
+
*Gel extraction(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>_XP)
-
*PCR amplification -> Digest(<nowiki>#</nowiki>29_EP, <nowiki>#</nowiki>20-3-29-5_EP, <nowiki>#</nowiki>3-29-5_EP)
+
*PCR amplification -> Digest(<nowiki>#</nowiki><html><span class="part_num">29</span></html>_EP, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_EP, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_EP)
-
*Digest(<nowiki>#</nowiki>20_SP)
+
*Digest(<nowiki>#</nowiki><html><span class="part_num">20</span></html>_SP)
*Sequencing prep.
*Sequencing prep.
*Ethanol precipitation (10ul TE)
*Ethanol precipitation (10ul TE)
-
**<nowiki>#</nowiki>3-11-5(pSB1C3): 144.5ng/uL
+
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>(pSB1C3): 144.5ng/uL
-
**<nowiki>#</nowiki>3-14-5: 728.0ng/uL
+
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>: 728.0ng/uL
-
**<nowiki>#</nowiki>3-17-5(pSB1C3): 505.7ng/uL
+
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>(pSB1C3): 505.7ng/uL
-
*Ligation(<nowiki>#</nowiki>31_EP-pSB1C3_EP, <nowiki>#</nowiki>36-3-29-5_ES-<nowiki>#</nowiki>20-3-37-5_XP-pSB1C3_EP)
+
*Ligation(<nowiki>#</nowiki><html><span class="part_num">31</span></html>_EP-pSB1C3_EP, <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_ES-<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>_XP-pSB1C3_EP)
*Luciferase assay
*Luciferase assay
*Asp diffusion assay
*Asp diffusion assay
Line 1,156: Line 1,159:
==='11/09/21(Wed)===
==='11/09/21(Wed)===
*Making plates
*Making plates
-
*Gel extraction(<nowiki>#</nowiki>20_SP, <nowiki>#</nowiki>29_EP, <nowiki>#</nowiki>3-29-5_EP, <nowiki>#</nowiki>20-3-29-5_EP)
+
*Gel extraction(<nowiki>#</nowiki><html><span class="part_num">20</span></html>_SP, <nowiki>#</nowiki><html><span class="part_num">29</span></html>_EP, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_EP, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_EP)
*Ligation
*Ligation
-
**<nowiki>#</nowiki>20_SP-<nowiki>#</nowiki>3-27-5_XP, <nowiki>#</nowiki>24_SP-<nowiki>#</nowiki>3-14-5-2-3-17-5_XP, <nowiki>#</nowiki>29_EP-pSB1C3_EP, <nowiki>#</nowiki>3-29-5_EP-pSB1C3_EP, <nowiki>#</nowiki>20-3-29-5_EP-pSB1C3_EP, <nowiki>#</nowiki>3-14-5-2-3-17-5_EP-pSB1C3_EP
+
**<nowiki>#</nowiki><html><span class="part_num">20</span></html>_SP-<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">24</span></html>_SP-<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_XP, <nowiki>#</nowiki><html><span class="part_num">29</span></html>_EP-pSB1C3_EP, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_EP-pSB1C3_EP, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_EP-pSB1C3_EP, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_EP-pSB1C3_EP
*Transformation
*Transformation
-
**<nowiki>#</nowiki>36 from BioBrick? part
+
**<nowiki>#</nowiki><html><span class="part_num">36</span></html> from BioBrick? part
-
**<nowiki>#</nowiki>33-3-14-5-2-3-17-5 into BL21 comp.
+
**<nowiki>#</nowiki><html><span class="part_num">33</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html> into BL21 comp.
-
*Digest(<nowiki>#</nowiki>3-14-5-2-3-17-5_SP, <nowiki>#</nowiki>3-14-5-2-3-17-5_ES, <nowiki>#</nowiki>3-29-5_ES)
+
*Digest(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_SP, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_ES, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_ES)
-
*Luciferase Assay(<nowiki>#</nowiki>36-3-14-5-2-3-17-5)
+
*Luciferase Assay(<nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
*Cell diffusion assay
*Cell diffusion assay
 +
<html></div></html>
 +
<html><div class="labDay"></html>
 +
==='11/09/22(Thu)===
 +
*Ligation (retry)
 +
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>_XP-<nowiki>#</nowiki><html><span class="part_num">20</span></html>_SP, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span>-</html><nowiki>#</nowiki><html><span class="part_num">24</span></html>_SP, <nowiki>#</nowiki><html><span class="part_num">29</span></html>_EP-pSB1C3_EP, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_EP-pSB1C3_EP, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_EP-pSB1C3_EP, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_EP-pSB1C3_EP
 +
*Colony PCR(<nowiki>#</nowiki><html><span class="part_num">36</span></html>(->culture for mini prep), <nowiki>#</nowiki><html><span class="part_num">33</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>(->Maybe the amplification was failed))
 +
*Gel extraction(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_SP, _ES; <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_ES)
 +
*Making plates
 +
**M9 0.25% Agar. +IPTG +Amp.: 20
 +
**LB Cm.: 20
 +
*Ligation
 +
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_SP-<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>_XP,
 +
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_ES-<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>_XP-pSB1C3,
 +
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_ES-<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>_XP-pSB1C3_EP
<html></div></html>
<html></div></html>
-
<html></div></html>  
+
<html><div class="labDay"></html>
 +
==='11/09/23(Fri)===
 +
*colony PCR
 +
*Column purification
 +
*Ligation
 +
*PCR amplification
 +
*Competent cell(BL21(DE3))
 +
*Asp diffusion test
 +
*Cell diffusion assay
 +
*Asp chemotaxis assay
 +
<html></div></html>
 +
<html><div class="labDay"></html>
 +
==='11/09/26(Mon)===
 +
*Ligation(<nowiki>#</nowiki><html><span class="part_num">20</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">24</span>-</html><nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, recNp-3-14-5-2-3-17-5)
 +
*Digest(pSB1C3_ES)
 +
*Miniprep(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span>-</html>pSB1C3, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span>-</html>pSB1C3, <nowiki>#</nowiki><html><span class="part_num">36</span></html>)
 +
*PCR amplification(pSB1A3, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>)
 +
*Ethanol precipitation(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span>-</html>pSB1C3, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span>-</html>pSB1C3)
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/09/27(Tue)===
 +
*Colony PCR(<nowiki>#</nowiki><html><span class="part_num">24</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">35</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
 +
*Digest(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_ES, <nowiki>#</nowiki><html><span class="part_num">36</span></html>_SP, pSB1C3_ES, pSB1C3_XP)
 +
*Ligation(<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>_XP-<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_SP, pSB1C3_ES, <nowiki>#</nowiki><html><span class="part_num">31</span></html>_EP-pSB1C3_EP)
 +
*Gel digestion(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_ES, <nowiki>#</nowiki><html><span class="part_num">36</span></html>_SP, pSB1C3_ES, pSB1C3_XP)
 +
*Nanodrop(<nowiki>#</nowiki><html><span class="part_num">36</span></html>_SP, pSB1C3)
 +
*PCR amplification(J04450)
 +
*Gel extraction(pSB1C3_ES)
 +
 +
<html></div></html>
 +
<html><div class="labDay"></html>
 +
==='11/09/28(Wed)===
 +
*Nanofrop
 +
**From PCR : <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>, pSB1C3
 +
**From Miniprep : <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>, J04450
 +
*Transformation(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">5</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>) -> BL21(DE) comp.
 +
*PCR amplification and Digest(EPcut) (<nowiki>#</nowiki><html><span class="part_num">31</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>)
 +
*Digest(pSB1C3_EP, pSB1C3_XP, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_ES, <nowiki>#</nowiki><html><span class="part_num">24</span></html>_SP_SP, <nowiki>#</nowiki><html><span class="part_num">24</span></html>_SP)
 +
*Sequencing(<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
 +
*Making AspA culture
 +
*Cell diffusion assay
 +
*Asp taxis assay
 +
*Gel extraction and Nanodrop(pSB1C3_EP, pSB1C3_XP, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_ES, <nowiki>#</nowiki><html><span class="part_num">24</span></html>_SP)
 +
*Ligation
 +
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_ES-<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>_XP-pSB1C3_EP
 +
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_XP-pSB1C3_XP
 +
**<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>_XP-pSB1C3_XP
 +
**<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>_XP-pSB1C3_XP
 +
**<nowiki>#</nowiki><html><span class="part_num">29</span></html>_XP-pSB1C3_XP
 +
**<nowiki>#</nowiki><html><span class="part_num">24</span></html>_SP-<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_XP
 +
**<nowiki>#</nowiki><html><span class="part_num">36</span></html>_SP-<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_XP
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/09/29(Thu)===
 +
*Ligation(<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>_EP-pSB1C3_EP)
 +
*Gel extraction and Nanodrop(<nowiki>#</nowiki><html><span class="part_num">31</span></html>_EP, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>_EP)
 +
*PCR amplification(pSB1C3, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
 +
*Digest(pSB1C3_EP, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_XP)
 +
*Colony PCR, PCR amplification, Miniprep and Colony diffusion assay(<nowiki>#</nowiki><html><span class="part_num">36</span></html>_SP-<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_XP)
 +
*Colony PCR and Miniprep(<nowiki>#</nowiki><html><span class="part_num">24</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/09/30(Fri)===
 +
*Digest(<nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_ES)
 +
*PCR amplification and Digest(EPcut)(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">29</span></html>, <nowiki>#</nowiki><html><span class="part_num">31</span></html>)
 +
*Digest(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_EP, <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_ES, pSB1C3_EP, pSB1C3_ES)
 +
*Culture for MIniprep from plate stock(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
 +
*Making competent cell(JM109)
 +
*Gel extraction and Nanodrop(<nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_ES_PCR, <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_ES_Miniprep, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_EP, pSB1C3_EP, pSB1C3_ES, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_EP, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>_EP, <nowiki>#</nowiki><html><span class="part_num">29</span></html>_EP, <nowiki>#</nowiki><html><span class="part_num">31</span></html>_EP)
 +
*Ligation(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_EP-pSB1C3_EP, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>_EP-pSB1C3_EP, <nowiki>#</nowiki><html><span class="part_num">29</span></html>_EP-pSB1C3_EP, <nowiki>#</nowiki><html><span class="part_num">31</span></html>_EP-pSB1C3_EP, <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>_ES-<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>_XP-pSB1C3_EP, recNp_ES-pSB1C3_ES, sulAp_ES-pSB1C3_ES, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_EP-pSB1C3_EP)
 +
*Sequencing prepartion(<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>_F, <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>
 +
*Miniprep, Digest(EXcut) and Nanodrop(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
 +
*Culture for Miniprep(pSB1C3, <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>)
 +
*Making plate LB Cm
 +
<html></div></html>
 +
<html></div></html>
 +
 +
<html><div class="labMonth" id="labMonth_10"></html>
 +
==October==
 +
<html><div class="labDay"></html>
 +
==='11/10/01(Sat)===
 +
*Part Sequencing for parts shipping
 +
*Miniprep(<nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>)
 +
*Sequencing preparation and Colony PCR(<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>_F, <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">24</span>-<span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
 +
*Colony PCR(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
 +
*second colony PCR
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/10/02(Sun)===
 +
*PCR amplification(using Quick Taq), Digest(Epcut), Gel extraction and Nanodrop(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>)
 +
*Gel extraction(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_EX)
 +
*Dual Luciferase assay(<nowiki>#</nowiki><html><span class="part_num">24</span></html>)
 +
*<nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html> Assay
 +
*<nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span>-<span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html> Assay
 +
*Ligation(recNp_ES-<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_EX, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>_EP-pSB1C3_EP)
 +
*Making agarose gel
 +
*Asp degradation assay
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/10/03(Mon)===
 +
*Miniprep, Nanodrop and PCR sequencing(<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">29</span></html>, <nowiki>#</nowiki><html><span class="part_num">31</span></html>, <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span>-<span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>, recNp, sulAp)
 +
*Colony PCR(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span>-</html>pSB1C3)
 +
*Preparation for shipping
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/10/04(Tue)===
 +
*Preparation for shipping(<nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">11</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">14</span>-<span class="part_num">5</span>-<span class="part_num">2</span>-<span class="part_num">3</span>-<span class="part_num">17</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">29</span></html>, <nowiki>#</nowiki><html><span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">36</span>-<span class="part_num">3</span>-<span class="part_num">29</span>-<span class="part_num">5</span>-<span class="part_num">20</span>-<span class="part_num">3</span>-<span class="part_num">37</span>-<span class="part_num">5</span></html>, <nowiki>#</nowiki><html><span class="part_num">31</span></html>, <nowiki>#</nowiki><html><span class="part_num">33</span></html>, <nowiki>#</nowiki><html><span class="part_num">35</span></html>)
 +
<html></div></html>
-
<!-- end of div#labMonth -->
+
<html></div></html>  <!-- end of div#labMonth -->
<html>
<html>
Line 1,244: Line 1,374:
   // part description popup
   // part description popup
-
  $('div#labnote *').each(function() {
 
-
    var $t = $(this);
 
-
    var txt = $t.html();
 
-
    //if ($t.children().length > 0) return;
 
-
    var expr = /#(\d+)(-(\d+))*/g;
 
-
    var subexpr = /(\d+)/g;
 
-
    var m;
 
-
    while (m = expr.exec(txt)) {
 
-
      var after = m[0].replace(/(\d+)/g, '<span class="part_num">$1</span>');
 
-
      txt = txt.replace(m[0], after);
 
-
    }
 
-
    $t.html(txt);
 
-
  });
 
   var description = {};
   var description = {};
   $('#partslist tr td:first-child').each(function() {
   $('#partslist tr td:first-child').each(function() {
Line 1,263: Line 1,380:
     var num = m[1];
     var num = m[1];
     var msg = '';
     var msg = '';
-
     var idx2name = { 0: 'Part', 1: 'Content', 2: 'Plasmid', 3: 'Length' };
+
     var idx2name = { 0: 'Registry ID', 1: 'Content', 2: 'Plasmid', 3: 'Length' };
     $(this).siblings().each(function(i) {
     $(this).siblings().each(function(i) {
       msg += idx2name[i] + ': ' + $(this).html() + '<br />';
       msg += idx2name[i] + ': ' + $(this).html() + '<br />';

Latest revision as of 20:25, 5 October 2011

Parts List

Each part is represented by s number (e.g. #20 for lac promotor) in this notebook. By pointing the mouse on the number within notebook entires, you can find out the details of the part.


Table 1. List of iGEM parts used in our project
Number Reg.ID Content Plasmid Length (bp)
#1 [http://partsregistry.org/wiki/index.php?title=Part:BBa_J23119 J23119] c.Promoter (Strong) pSB1A2 35
#2 [http://partsregistry.org/wiki/index.php?title=Part:BBa_J23118 J23118] c.Promoter (Medium) [http://partsregistry.org/wiki/index.php?title=Part:BBa_J61002 BBa_J61002] 35
#3 [http://partsregistry.org/wiki/index.php?title=Part:BBa_B0032 B0032] RBS pSB1A2 13
#5 [http://partsregistry.org/wiki/index.php?title=Part:BBa_B0014 B0014] d.Ter pSB1AK3 95
#9 [http://partsregistry.org/wiki/index.php?title=Part:BBa_E0240 E0240] RBS-GFP-d.Ter pSB1A2 876
#10 [http://partsregistry.org/wiki/index.php?title=Part:BBa_E0040 E0040] GFP pSB1A2 681
#11 [http://partsregistry.org/wiki/index.php?title=Part:BBa_E1010 E1010] RFP pSB2K3 723
#14 [http://partsregistry.org/wiki/index.php?title=Part:BBa_I712019 I712019] fLuc pSB1AK8 1653
#17 [http://partsregistry.org/wiki/index.php?title=Part:BBa_J52008 J52008] rLuc pSB1AK3 936
#20 [http://partsregistry.org/wiki/index.php?title=Part:BBa_R0011 R0011] lacP pSB1A2 55
#21 [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0012 C0012] LacI pSB1A2 1128
#22 [http://partsregistry.org/wiki/index.php?title=Part:BBa_I712074 I712074] pT7 pSB1AK8 46
#23 [http://partsregistry.org/wiki/index.php?title=Part:BBa_K145001 K145001] T7 RNA Pol. pSB1A2 2655
#24 [http://partsregistry.org/wiki/index.php?title=Part:BBa_J22106 J22106] recAp pSB1A2 192
#27 [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0083 C0083] AspA pSB2K3 1518
#28 [http://partsregistry.org/wiki/index.php?title=Part:BBa_K112808 K112808] T4 phage lysis device (no promoter) pSB1A2 1785
#29 - CheZ pSB1AK3 728
#30 [http://partsregistry.org/wiki/index.php?title=Part:BBa_K117000 K117000] Lysis gene pSB1A2 144
#31 - LexA pSB1AK3 750
#33 - sulAp pSB1AK3 67
#34 - uvrAp pSB1AK3 96
#35 - recNp
#36 [http://partsregistry.org/wiki/index.php?title=Part:BBa_R0051 R0051] cI-repressed promoter pSB1A2 49
#37 [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0051 C0051] cI repressor (LVA tagged) pSB1A2 \
#38 - RecA

Lab Notebook

  • May
  • June
  • July
  • August
  • September
  • October

Please enable Javascript to view this calendar.

May

'11/05/18 (Wed)

  • Making LB medium, 50×TAE, Tris-HCl (pH8.0)

'11/05/24 (Tue)

  • Making SOB medium, 0.5M EDTA (pH8.0)

'11/05/25(Wed)

  • Making TB(pH=6.7), LB plate

'11/05/26(Thu)

  • Making Mgaq(for SOB medium), Competent cell
  • TB filtration

'11/05/31(Tue)

  • iGEM parts resuspension + frozen stock (at -20℃)
  • Transformation
  • Overnight culture on LB plate (with 100ug/ml ampicilline)

June

'11/06/01(Wed)

  • Picking up colony and transfer to LB medium
  • Making LB medium

'11/06/02(Thu)

  • Miniprep
  • Making Glycerol(50%)(for cryopreservation)

'11/06/07(Tue)

  • Nanodrop
  • Transformation
  • Culture from frozen stock

'11/06/08(Wed)

  • Picking up colony

'11/06/09(Thu)

  • Miniprep

'11/06/13(Mon)

  • Planting Negative control
  • Making frozen stock
  • Transformation(BBa_E0240, #3, B0015, #5, #10, ,J31000, ,J44000)

'11/06/14(Tue)

  • Picking up colony
  • Making Mg reagent

'11/06/15(Wed)

  • Miniprep

'11/06/17(Fri)

  • Picking up colony(#3, B0015, B0040,#9, J31000, J44000)
  • Miniprep(#5)
  • Dissolution(#1, #2, K16500, #24)

'11/06/21(Tue)

  • Making frozen stock(#3, B0015, B0040, #9, J31000, J44000)
  • Passafe culture(#9, J31000)
  • Nanodrop again

'11/06/22(Wed)

  • Miniprep(#9, J31000)
  • Making competent cell

'11/06/24(Fri)

  • Digest (product of Miniprep 06/09 EScut)
  • Making agarose gel
  • Electrophoresis

'11/06/28(Tue)

  • Digest(product of Miniprep 06/09 EPcut)
  • Electrophoresis
  • Defrost and transformation(BBa_E0030, E0020, #14, I712052,#17
  • Making 1×TAE, agarose gel, LB medium, LB plate(amp),

'11/06/29(Wed)

  • Picking up colony
  • Making LB broth

July

'11/07/01(Fri)

  • Digest(EPcut)
  • Miniprep(BBa_E0030, E0020, #14, I712052, #17)

'11/07/05(Tue)

  • Electrophoresis(product of digest 07/01)
  • Digest(#9, #14, #17, #3, #5)
  • Making antibiotic stock(1000×)(Km,Cm)

'11/07/06(Wed)

  • Gel extraction (pre)(BBa_I712052)
  • Picking up colony(Ba_J23119, #2, #24, #10)
  • Making agarose gel

'11/07/07(Thu)

  • Digest(BBa_E0240, #14, #17, #3, #5
  • Defrost(BBa_E1010, K325101, #20, #21, #22, #23
  • Transformation(BBa_K145001, #20, #21, #22
  • Making LB plate(Cm,Km)

'11/07/08(Fri)

  • Miniprep(BBa_J23119, #2, #10, #24)

'11/07/11(Mon)

  • Gel extraction(#9 #14, #17)
  • Transformation(#1, #11, #20, #21, K325101, #22, #23)

'11/07/12(Tue)

  • Picking up colony

'11/07/13(Wed)

  • Frozen stock(#1(18A), #20(6G), #21(2O), #14(6N), #23(2F))
  • Picking up colony(#11)
  • Gel extraction(B#3, #5)
  • Digest (#2 SPcut)

'11/07/14(Thu)

  • Miniprep(#1, #11, #20, #21, #22, #23)
  • Electrophoresis(#2)
  • Gel extraction(#2, #3, #5)
  • Passafe(#11)
  • Making LB+amp

'11/07/15(Fri)

  • Ligation(#2-#9, #2-#3, #14-#5, #17-#5)
  • Transformation(#2-#9, #2-#3, #14-#5, #17-#5)
  • Dispensing Ligation Buffer
  • Frozen stock(#11)

'11/07/19(Tue)

  • Digest
  • Gel extraction
  • Making competent cell

'11/07/20(Wed)

  • Picking up colony
  • Making Master plate
  • Testing product of Miniprep
  • Transformation (#2, #27, #28)

'11/07/21(Thu)

  • Miniprep(#2-3, #2-9, #14-5, #17-5, #11, #20)
  • Testing product
  • Digest
    • SPcut : #20, #2-3
    • XPcut : #10, #11, #14-5, #17-5
  • Making TB

'11/07/22(Fri)

  • Frozen stock(#2, #2-3, #2-9, #14-5, #17-5
  • Electrophoresis(#20_SP, #2-3_SP, #10_XP, #11_XP, #14-5_XP, #17-5_XP)
  • Digest
    • SPcut : #20
    • XPcut : #10, #11, #14-5, #17-5
  • Transformation(#27, #28, product of Miniprep 07/20)

'11/07/25(Mon)

  • Colony check(#2, #2-9)
  • Gel Extraction(#20_SP, #10_XP, #11_XP, #14-5_XP, #17-5_XP)
  • Ligation and Transformation(#20-#9, #3-#11, #3-#14-5, #3-#17-5)
  • Defrost Primer(200×, 10×)
  • Dispensing PCR Mix

'11/07/26(Tue)

  • Picking up colony and making master plate(#20-9, #3-11, #3-14-5, #3-17-5)
  • Colony PCR(#20-9, #3-11, #3-14-5, #3-17-5)
  • Digest(#14-5 XPcut,Ecut, #22 SPcut, #23 XPcut)
  • Ligation(#3-#14-5(Re), #3(Negative control))
  • Transformation(#15, #27, #28, #30, #3-14-5(Re), #3(Negative control))
  • Making reagent for TB(KOH, CaCl2, KCl, MnCl2)

'11/07/27(Wed)

  • Miniprep(#1, #2, #3, #10, #22, #24, #20-9, #3-11, #3-17-5)
  • Electrophoresis

'11/07/28(Thu)

  • Gel extraction(#14-5_XP, #22_SP, #23_XP)
  • Digest
    • SPcut : #1,#2, #3, #24
    • EScut : #3-11
    • XPcut : #3-17-5
  • Ligation(#3-#14-5)
  • Making TB bugger, LB plate, 0.3% LB plate

'11/07/29(Fri)

  • Cloning(lexA, cheZ)
  • Colony PCR(#3-#14-5)
  • Gel extraction(#1_SP, #2_SP, #3_SP, #24_SP, #3-11_ES, #3-17-5_XP, lexA, cheZ PCR product)
  • Miniprep(#30)
  • Digest(lexA, cheZ PCR product XP cut)

August

'11/08/01(Mon)

  • Nanodrop(#3-11(1), #1, #3, #24, #2, #3-17-5(4)M#30)
  • Digest(#3 SPcut, #5 EXcut, #9 XPcut, #30 XP cut)
  • Cloning(lexA, cheZ)
  • Gel extraction(#3, #9, #30, pSB1A2)
  • Ligation(#2-#3-17-5, #3-11-#5, #14-#5, #3-#23, #22-#9, #3-#30)
  • Transformation(#2-#3-17-5, #3-11-#5, #14-#5, #3-#23, #22-#9, #3-#30, #27(IPTG), #28)

'11/08/02(Tue)

  • Colony PCR(#2-#3-17-5, #3-11-#5, #14-#5, #3-#23, #22-#9)
  • Cloning(PCR)(#27, #28)
  • Gel extraction
  • PCR_2nd(lexA, cheZ, #27, #28)
  • Digest (#30 again)
  • Making competent cell(cheZ-, Wild Type)
  • Miniprep

'11/08/04(Thu)

  • Electrophoresis(product of ligation #2-3-17-5(2), #3-11-5(1,2), #14-5(2), #22-9(1,2,3))
  • Frozen stock(#2-3-17-5(2), #3-11-5(1,2), #14-5(2), #22-9(1,2))
  • Making Gel
  • Digest(#23 XPcut)
  • IPTG induct(#20-9)

'11/08/05(Fri)

  • Digest(#14-5(2) XPcut, #3-11-5(1) XPcut)
  • Cloning(PCR)(#27, #28, lexA, cheZ)

'11/08/08(Mon)

  • Defrost Frozen stock, Miniprep, Nanodrop and Electrophoresis(#5, #20, #23, #30)
  • Making Gel
  • Digest
    • XPcut : #27, #28, cheZ, lexA, #3-11-5(1), #23,#30
    • EScut : #6
    • EXcut : #7
    • SPcut : #7,#20
  • Defrost Frozen stock(#5, #14-5(2), #3-11-5(1))

'11/08/09(Tue)

  • Miniprep, Electrophoresis and Digest(#5, #14-5(2), #3-11-5(1))
  • Gel extraction and Nanodrop(#6, #27, #8, cheZ, lexA③, #3-11-5(1), #23, #7_EX, #20, #7_SP)
  • Ligation(#27-psB1A2, #3-27, #20-28, #28-psB1A2, #3-29, #29-psB1A2, #31-psB1A2, #3-#23, #1-#3-11-5(1), #2-#3-11-5(1), #24-#3-11-5(1), #3-#30)
  • Making LB plate

'11/08/10(Wed)

  • Colony PCR(#27-pSB1A2, #3-27, #20-28, #28-psB1A2, #3-29, #29-psB1A2, #31-pSB1A2, #3-#23, #1-#3-11-5(1), #2-#3-11-5(1), #24-#3-11-5(1), #3-#30))
  • Cloning(lexA, cheZ)
  • Digest
    • EScut : #6
    • EXcut : #7
    • SPcut : #1, #2, #3, #7, #20
    • XPcut : #23
  • Defrost Primer

'11/08/11(Thu)

  • Gel extraction(#30, #14-5(insert,vector), #5, #3-11-5, cheZ, lexA)
  • Cloning(sulAp, uvrAp)
  • Miniprep(#3, #20-28, #31, #24-3-11-5, #3-30)

'11/08/12(Fri)

  • Colony PCR(#3-14-5(2), #6-5, #3-23)
  • Cloning(nested-PCR)(uvrAp, sulAp)
  • Check Electrophoresis
    • 8/11 PCR product:#27, #28
    • Miniprep product:#3-14-5(2), #5
  • Digest(#5 EXcut, #14 EScut)
  • Gel extraction
    • 8/11 digest product : #5, #14-5, #3-30, cheZ, lexA
    • 8/11 PCR product : #27, #28
    • 8/12 nested-PCR product : sulAp, uvrAp
  • Colony PCR (re) (#13-14-5)
  • Miniprep(ligation product : #6-5, #3-23)
  • Cloning(hotstart-PCR: uvrBp , cheZ)
  • Digest
    • XPcut:#27, #28, #6-5
    • EScut: sulAp, uvrAp, #3-23

'11/08/15(Mon)

  • Gel extraction(from previous digest products(8/12) : #27, #28, #33, #34, #4, #3-23)
  • Digest
    • XPcut: cheZ, uvrBp(PCR products)
    • EScut: #3-11-5 (to obtain pSB1AK3 EScut)
  • Assay of Cell lysis construction(pLuc-Lysis device) inducted by IPTG
  • Making Lysis buffer, Agarose gel
  • Gel extraction(from digest products XPcut: cheZ, uvrBp(PCR products) EScut: #3-11-5 to obtain pSB1AK3 EScut)
  • Ligation(#3-#14-5, #24-#3-17-5, #24-#3-11-5, #20-#28, #3-#27, #3-#29, #3-23-#5, #33-pSB1AK3, #34--pSB1AK3, #35--pSB1AK3)
  • Transformation(ligation products:WT, #2-9:cheZ-, #14, #17) -> 37 incubation

'11/08/16(Tue)

  • Colony PCR(#3-#14-5, #24-#3-17-5, #20-#28, #3-#27, #3-#29, #3-23-#5)
  • Culture (#20-#28, #5, #14, #17, #2-9, #2-4, #2-3-17-5, #1, #2)
  • Incubation (#24-#3-11-5, #33-pSB1AK3, #34-pSB1AK3, #35-pSB1AK3)
  • Colony PCR(additional)(previously incubated ligation products: 3, 8, 10)
  • hotstart PCR(sulAp, uvrAp, uvrBp)
  • Digest(SPcut: #3, Ecut:#3(control)) go to 37 incubation
  • Ligation retry(#3-#14-5, #3-#27, #3-#29, Negative control :#3)
  • Frozen stock(#14, #17)
  • Culture test(#14 on 0.3%Agar.(km))
  • Ethanol precipitation: 29.9ng/ul * 1000ul
  • Miniprep(Ligation products : #2-3-17-5, #14, #17, #1, #2)
  • Electrophoresis(Colony PCR products: 3, 8, 10)
  • Waking from frozen stock(#3, CheZ-, WT)

'11/08/17(Wed)

  • Hotstart-PCR (Change conditions a little)
    • From Genome
    • From nested-region
    • From parts
  • Making Competent Cell, SOB medium
  • Ethanol precipitation(WT: 40ng/ul)
  • Culture from master-plate(#6-5, #20-28, #3-27, #33, #24-3-11-5 #24-3-17-5) -> Miniprep
  • Assay(IPTG induced cell lysis device
  • Gel extraction(SPcut:#3)
  • Culture from Frozen stock(#1, #2, #2-3-17-5)
  • Electrophresis check(Loading only DNA size markers)
  • Miniprep(#3, #6-5, #20-28, #3-27, #33, #24-3-11-5 #24-3-17-5)

'11/08/18(Thu)

  • Digest(8/17#3, #33 SPcut, #14-5 XPcut, #33 Ecut, #3-27,#3-30,#20-28 EScut, pSB1K3 EPcut)
  • Hotstart-PCR (Change conditions a little)
    • From Genome : uvrBp, cheZ
    • From nested-region : uvrA
  • Culture from 0809 master-plate ⑪ (#2-3-11-5) → Miniprep.
  • Making LB plate & tube with various agarose conc. (plate: 0.5, 0.7, 0.9%×2 each, tube: 0.5,1.0,1.5,2.0,3.0%×2 each)(selection: Kanamycin)
  • Gel extraction(8/17#3_SP,#33_SP, #14-5_XP, #33_E, #3-27_ES, pSB1K3 _EP)
  • Miniprep(#1, #2, #2-3-11-5, #2-3-17-5)
  • Digest(#1,#2 SPcut, #2-3-17-5 XPcut, #2-3-11-5 SPcut(check), EPcut(selection), #3-30, #20-28 EScut, 0817 Gel extracted PCR product (uvrBp) EScut, 0818PCR products (uvrAp, uvrBp, cheZ) ES&XPcut)
  • Ligation (8/17#3-#14-5, 8/17#3-8/15#29, #33-#3-11-5, 8/15#34-pSB1AK3, #3-27-#5, #20-28-pSB1AK3, #28-pSB1AK3)
  • Culture for frozen stock in eppendolf tube

'11/08/19(Fri)

  • ColonyPCR(from ligation products)(#3-14-5, #3-29, #33-3-11-5, #34-pSB1AK3, #3-27-5)
  • Gel extraction(08/18's digest products:#1_SP, #2_SP, #2-3-11-5_SP, #2-3-17-5_XP, cheZ_XP, #2-3-11-5_EP, 8/18uvrBp_ES, 8/17uvrBp_ES, uvrAp_ES)
  • Colony PCR retry(#3-29, #34-pSB1AK3)
  • Frozen Stock
  • Check the list of frozen stock
  • Assay of recAp from biobrick parts using RFP construct & transilluminator
  • Electrophoresis(#3, #3-27, #3-30, #6-5, #7, #20-28, #24-3-11-5, #24-3-17-5, #33)

'11/08/22(Mon)

  • Making SOB medium
  • Culture for Miniprep
    • From frozen stock(#1, #2, #7, #2-3-17-5(2), #3-30)
    • From master plate(#3-14-5, #33-3-11-5, #34-pSB1AK3, #3-27-5)
    • From master plate retry( at 13:50 OD600=0)
    • From ligation products (for mistake above)
  • Miniprep, Nanodrop and Electrophoresis
    • From frozen stock(#1, #2, #2-3-17-5(2), #3-30) #7 was a little bit weak
    • From ligation products(#7, #3-14-5, #3-27-5, #34, #33-3-11-5)
  • Digest
    • From Frozen Stock (#7_EX, #30_XP, #2-3-17-5_XP, #3-30_ES)
    • From Master plate(#3-14-5_XP, #34-pSB1AK3_SP, #3-27-5_XP)
  • Sulvage #33 using protocol of transformation
  • Culture for Frozen stock
    • From Master plate(#24-3-17-5, #14-5), 700ul, in eppendolf tube

'11/08/23(Tue)

  • Gel extraction
    • from O.N. digest products(#7_EX, #30_XP, #2-3-17-5_XP, #3-30_ES, #3-14-5_XP, #34-pSB1AK3_SP, #3-27-5_XP)
  • Frozen stock
    • From O.N. culture in eppendolf tube 700ul(#24-3-17-5, #14-5)
    • From O.N. culture(for Miniprep)(#34, #3-27-5, #33-3-11-5)
  • Miniprep
    • retry from master plate #3-14-5(278.1 ng/ul), #33-3-11-5(188.9ng/ul), #34-pSB1AK3(89.6ng/ul), #3-27-5(327.0 ng/ul) O.N. culture
  • Digest (#14-5 XPcut, #3-27-5 Xcut, Pcut (check), #3-27-5 EScut (sulvage) )
  • Ligation (#14-#5, #3-30-#5, #3-#30, #34-#3-11-5 #20-#28)
  • #33 sulvage (isolation culture)
  • Making culture(LB amp, LB)

'11/08/24(Wed)

  • Gel extraction & Electrophoresis(#3-27-5_ES, #14-5_XP, #3-27-5_X, *PCR for amplification (or cloning) -> PCR clean-up system -> digest
    • #27 (82.1 ng/ul, total = 30 ul) -> XP cut
    • #28 (104.4 ng/ul, total = 30 ul) -> XP cut
  • Making plate and amp stock ->4℃
    • amp 32
    • km 11 (km conc. = 3/4 * that it was)
  • Miniprep and Nanodrop
    • #33 32.1 ng/ul <- there is probability that culture volume was not enough ->transfromation
  • Luciferase assay reagents prep
    • D-Luciferin Solution : 1ml*10 + 100ul*10 at -80℃
    • Coelenterazine Solution : 100ul * 10 at -20℃
    • Lysis buffer : 500ul * 10 at -20℃
  • Ligation(#3-30-#5, #3-11-5-#34, #30-#3, #28-#20, #35-pSB1AK3, #2-3-11-5-pSB1K3, #14-5-#3 and Negative control)

'11/08/25(Thu)

  • Renilla luciferase assay (with #2-3-17-5 culture)
  • Gel extraction (#27, #28 XPcut)
  • Ligation (0824 retry & #20-#28, #3-#27)
  • Transformation (Ligation products into WT competent cells & #14-5 miniprep product into WT cell (line-drawing spread), #33 miniprep product into ΔcheZ cell (line-drawing spread))
  • Digest (#14 XPcut)
  • Passage(#2-3-17-5, WT, ΔcheZ, WT in eppen, ΔcheZ in eppen)

'11/08/26(Fri)

  • Renilla luciferase assay (with #2-3-17-5 culture)
  • Colony PCR (#33 Miniprep transformations)
  • Gel extraction (#14_XP)
  • Ligation (retry) (using today's competent cells, 0802 ΔcheZ cells)
  • Cloning (cheZ)
  • Digest (#5 EXcut, cheZ PCR products XPcut)
  • Passage in a shaker(#2-3-17-5, WT, ΔcheZ)
  • Making competent cell, LB broth
  • Contamination check (0817 comp., 0802 comp., 0802 ⊿cheZ comp. spread on LB (amp) plates)

'11/08/27(Sat)

  • Ligation & Transformation (0826 ①②③⑤⑧⑨) (into Nippon Gene comp.)
  • Contamination check (WT FS, ΔcheZ Frozen stock)
  • Waking from Frozen stock(WT, ΔcheZ)

'11/08/28(Sun)

  • Making ΔcheZ comp., LB amp plates, TB
  • Colony PCR & master plate preparation -> Failure!!
  • Gel extraction (#29_XP, #5_EX)
  • Miniprep and frozen stocks (#33, #14-5)
  • Digest (#20 SPcut, #14-5 XPcut)
  • Waking from frozen stock (WT, ⊿cheZ, #20, #14-5)

'11/08/29(Mon)

  • PCR purification -> digest with XP(#27, #28)
  • Miniprep(#20, #14-5)
  • Gel extraction(#14-5, #20)
  • Ligation (higher I/V ratio, into cheZ comp.)(#3-#14-5(retry), #2-3-11-5 to pSB1K3 (retry), #35-pSB1AK3, (#33,#34)-#3-17-5)

'11/08/30(Tue)

  • Making LB amp, Aspartic acid solution (10mM), H2O2 solution (10mM)
  • Gel extraction -> PCR was failed!(#27,#28)
  • There was trash (from PCR) below
  • Colony PCR(#3-#14-5, #2-3-11-5 to pSB1K3, #35-pSB1AK3, (#33,#34)-#3-17-5)
  • PCR -> check (5uL EP -> PCR clean-up(#27,#28)
  • Digest(#20_SP, pSB1AK3_EP, #3_SP)
  • EP for MP check(#14-5, #20)
  • Ligation(#3-#29, pSB1AK3-#2-3-11-5 on Amp/Km plate, #3 (Negative control), pSB1AK3 (Negative control) on Amp/Km plate
  • Transformation(#2-9 into cheZ-) -> Line-drawing plating
  • SOS stress (UV irradiation or H2O2)(#24-3-11-5, #33-3-11-5)
  • culture for Miniprep from master plate(#34-3-17-5)
  • Waking from frozen stock(#24-3-11-5, #33-3-11-5)

'11/08/31(Wed)

  • Making LB plates(amp, km)
  • Luciferin reagent preparation
  • Flask A.C. for making competent cells
  • Miniprep (#34-3-17-5)
  • Gel extraction(#20_SP, pSB1AK3_EP, #3_SP)
  • PCR -> clean-up -> XPcut(#27, #28, #14-5, #3-30)
  • Waking from frozen stock(#2-9 (WT), #24-3-17-5)
  • Plating from liquid culture(#2-3-17-5)
  • Ligation(#20-#3-17-5, #2-3-11-5-pSB1AK3, #3-#14, #20-#28, #3-#27, #3-#29, #35-pSB1AK3, #3-#14-5, #3-30-#5, #33-#3-17-5 & Negative control)
  • Transformation
    • Plasmid isolation(#34-3-17-5(MP;1:9gradient))
    • Part cloning(#36, #37)
    • Contamination check(pSB1AK3(EPcut), pSB1K3(EPcut))
    • Titer check(#3-17-5(Takara comp.,1:9gradient), #2-3-11-5(ΔcheZ comp.0802, 1:9gradient), #2-3-11-5(ΔcheZ comp.0830, 1:9gradient))
    • Ligation products

September

'11/09/01(Thu)

  • Colony PCR
    • #34-3-17-5(MP), #3-14, #3-29, #35-pSB1AK3, #3-14-5, #37(from Part cloning)
  • Gel extraction
    • #27_XP ,#28_XP(0830); #27_XP, #28_XP, #3-30_XP -> failed!, #14-5_XP(0831)
  • Making medium(SOB, LB amp plate), competent cells (WT & ΔcheZ)
  • Miniprep(#3, #20)
  • PCR and Column purification
    • #3-14, #3-29, #3-14-5 (Recovery from colony PCR tubes!)
  • Digest
    • #3 -> Failed (Mixed with DMSO!), #20
    • #3-14, #3-29, #3-14-5
  • Ligation
    • #3-27, #20-#3-17-5
  • Transformation
    • #36 into Nippon Gene comp.
    • Ligation products into Takara comp. (including N.C.)
    • Titer check(MP product #3 into 0901 WT comp., MP product #20 into 0901 ΔcheZ comp.)
  • Mobility assay(#14-5 (km resistance) WT/ΔcheZ cells)

'11/09/02(Fri)

  • Making medium and reagent(LB 120ml(0.125, 0.25, 0.5% Agar, M9 stock(20×), M9 200ml, Vitamins Solution (100×))
  • Gel extraction -> Digest products were OK but extraction was failed.
    • #20_SP, #3-14_XP, #3-14-5_XP, #3-29_XP
  • Colony PCR & Master plate(#20-3-17-5, #3-27)
  • Miniprep(#3, #20, #37, #3-14, #3-29, #3-14-5, #20-3-17-5, #3-27, ΔcheZ)
  • Hot-start PCR & Column purification -> PCR products were OK but all purified products were lost.
    • #20-3-17-5, #3-27 (Recovery from colony PCR tubes)
    • #3-14, #3-14-5, #3-29 (from miniprep products)
  • PCR & Column purification
    • #20-3-17-5, #3-27, #3-14-5, #3-29 (from miniprep products)
  • Digest
    • #3_SP, #20_SP, #37_ES, #3-14_ES, #3-29_ES, #3-14-5_XP, ΔcheZ_E/P (from MP products)
    • #20-3-17-5_XP, #3-27_ES, #3-14-5_XP, #3-29_ES (from PCR products)

'11/09/03(Sat)

  • Gel extraction
  • Ligation & Transformation
    • #20-#3-14-5, #2-#3-14-5, #3-27-#5 into Nippon Gene comp. (+ High Competent Broth 240ul)
    • #3-27-#5, #37-#5 into 0902 WT comp.(+ Takara SOC broth 500ul)
  • Assays
    • Mobility assay (.5%, .25%, .125%)
    • Asp assay
  • Part Cloning (#36)

'11/09/04(Sun)

  • Hot-start PCR Pre-Mix (4×) preparation (Buffer + dNTPs + THUNDER Taq)
  • Colony PCR(#20-3-14-5, #3-27-5, #3-29-5, #37-5)
  • Colony Diffusion Assays
  • GFP Live-Imaging Assays
  • Ligation & Transformation (retry)
    • #2-#3-14-5, #3-27-#5, #3-29-#5, #37-#5, #20-#28 into 0901 WT comp.
    • #36 into Takara comp.
    • #2, #5 as Negative Controls into 0901 WT comp.
  • Culture for Miniprep(#20-3-14-5, #20)
  • Digest(#20)

'11/09/05(Mon)

  • Colony Diffusion Assays (#2-9) WT,cheZ-
  • Gel extraction(#20)
  • Luciferase Assays
    • Calibrations (Firefly Luciferase standards)
    • Background Assay
    • Dual Luciferase Assay(#20-3-14-5, #20-3-17-5)
  • Colony PCR(#2-#3-14-5, #3-27-#5, #3-29-#5, #37-#5, #20-#28, #36)
  • PCR -> clean-up -> Digest(#3-29-#5, #37-5)
  • IPTG-inducted Cell lysis test(IPTG+, LB plate (Positive control))
  • Miniprep(#20-3-14-5, #37-5, #36)
  • Making M9 -> Asp -> all failed(.125%, .25% Agar)
  • Culture for Miniprep(#2-3-14-5)

'11/09/06(Tue)

  • Gel extraction(#37-5, #3-29-5)
  • Miniprep(#2-3-14-5, #20-3-14-5, #20-3-17-5, #36, #3-29-5, #37-5)
  • PCR (retry) (Hot-start & Normal PCR) -> Purification -> Digest(#3-29-5_XP, #37-5_ES)
  • PCR (cloning retry)(lexA, uvrBp)
  • Ligation & Transformation
    • #3-27-#5, #3-30-#5, #2-3-11-5-pSB1AK3
    • #5, pSB1AK3 (N.C.)
  • Culture for GFP live-imaging (#2-9 WT/cheZ-) -> 4℃ storage
  • Ninhydrin sol. preparation (Not dissolvable!) -> 4℃ storage

'11/09/07(Wed)

  • Colony PCR(#3-27-5, #3-30-5)
  • Digest(#36_SP)
  • Culture from master plate(#36, #3-29-5, #37-5)
  • Miniprep(#36, #3-29-5, #37-5, #3-27-5 (failed))
  • PCR purification & Digest(lexA_XP, uvrBp_ES)
  • Gel extraction(#3-29-5_XP, #37-5_ES, pSB1AK3_EP, #36_SP)
  • PCR amplification -> Digest(#3-14-5, #3-29-5, #37-5, #3-27-5)
  • Digest(#36_SP (re), #2_SP, #2-3-17-5_EX, #3-14-5_ES, #34_SP)
  • Culture for Miniprep(#2, #34, #36, #3-27-5, #3-14-5, #2-3-17-5)

Ligation & Transformation

    • #33-#3-14-5, #34-#3-14-5, #36-#3-14-5, #2-#3-29-5, #20-#3-29-5, #3-#37-5, #2-3-11-5-pSB1AK3, #1-#3-17-5
    • #33, #34, #36, #1, #2, #20, #3, pSB1AK3 (N.C.)
  • Ninhydrin stock solution preparation
    • Asp conc. check (it took 5 min at 80C)
  • Making M9 medium(.25, .125% Agar.)

'11/09/08(Thu)

  • Making competent cell (WT), LB(amp) plate
  • Frozen stocks(#36, #3-14-5, #3-27-5)
  • Colony PCR(#33-3-14-5, #34-3-14-5, #36-3-14-5, #20-3-29-5, #2-3-11-5(psB1AK3))
  • Miniprep(#2, #34, #36, #2-3-17-5, #3-27-5, #3-14-5)
  • Gel extraction
    • PCR product: #35_SP, #31_XP, #3-14-5_XP, #3-27-5_XP, #3-29-5_XP, #37-5_XP
    • Miniprep product: #2_SP, #34_SP, #36_SP, #2-3-17-5_EX, #3-14-5_ES
  • PCR amplification(#34-3-14-5, #36-3-14-5, #20-3-29-5, #20-3-14-5)
  • PCR for Sequencing -> Store at -20C(#33-3-11-5, #34-3-14-5, #20-3-29-5, #37-5)
  • Ligation(#35_ES-psB1AK3_ES, #31_XP-psB1A2_XP, #31_XP-psB1AK3_XP, #3-#37-5, #36-3-29-5, #3-14-5_ES-#2-3-17-5_EX, #20-#3-27-5)
  • Culture for Miniprep(#34-3-17-5, #36-3-14-5, #20-3-29-5, #2-3-11-5)
  • Digest
    • PCR products: #34-3-14-5_ES, #36-3-14-5_ES, #20-3-14-5_ES
    • pSB1C3_EP
  • Asp Gradient test

'11/09/09(Fri)

  • Making competent cell (WT), TB
  • Colony PCR(#35-pSB1AK3 (failed), #31-pSB1AK3, #3-#37-5, #36-#3-29-5, #3-14-5-#2-3-17-5, #20-3-27-5(failed))
  • PCR amplification
    • From Miniprep : #3-14-5, #3-17-5
    • From Colony PCR : #31, #3-37-5, #36-3-29-5, #3-14-5-2-3-17-5 (failed)
  • Digest
    • Miniprep product: #3-11-5_EP
    • PCR-purified product: #3-14-5_EP, #3-17-5_EP, #31_EP, #3-37-5_XP, #36-3-29-5_ES
  • Miniprep(#34-3-14-5, #36-3-14-5, #20-3-29-5, #2-3-11-5 (pSB1AK3))
  • Gel extraction(#34-3-14-5_ES, #36-3-14-5_ES, #20-3-14-5_ES, #3-11-5_EP)
  • Ligation
    • #3-11-5_EP-pSB1C3_EP, #29_XP-pSB1AK3_XP
    • #36-3-14-5_ES-#2-3-17-5_EX, #20_SP-#3-27-5_XP(retry)
  • Comp. check
    • #2-3-11-5 (pSB1AK3) into 0908 WT comp.
    • #20-3-29-5 into 0909 WT comp.
    • #20-3-29-5 into 0802 cheZ- comp.
  • Part cloning(BBa_J04450 (IPTG-induced RFP reporter on pSB1C3))
  • Culture for Miniprep(#31-pSB1AK3, #3-37-5, #36-3-29-5, #3-14-5-2-3-17-5)
  • UV irradiation -> Luciferase Assay(#24-3-17-5, #34-3-17-5)
  • Asp Gradient test
  • CleanSEQ -> Sequencing(#33-3-11-5, #34-3-14-5, #20-3-29-5, #37-5)

'11/09/10(Sat)

  • Culture plate storage (4℃)
  • Miniprep(#31-pSB1AK3, #3-37-5, #36-3-29-5, #3-14-5-2-3-17-5)
  • Gel extraction(#3-14-5_EP, #3-17-5_EP, #31_EP, #3-37-5_XP, #36-3-29-5_ES)
  • Colony PCR(#36-3-14-5-#2-3-17-5 (Maybe Ligation was FAILURE , estimated length is 1000-1500 by electrophoresis))
  • Ligation(#20_SP-#3-37-5_XP)

'11/09/11(Sun)

  • Digest from Miniprep products
    • #3-14-5-2-3-17-5_XP
    • #36-3-14-5_SP, #20_SP -> O.N.
  • Colony PCR
    • #3-11-5 (pSB1C3), #29 (pSB1AK3), BBa_J04450 (pSB1C3)
    • #20-#3-27-5(failed), #20-#3-37-5, #36-#3-14-5-2-3-17-5
  • Gel extraction(#3-14-5-2-3-17-5_XP)
  • Culture for Miniprep
    • From Master plate : #20-3-37-5, #29 (pSB1AK3), BBa_J04450 (pSB1C3)
    • From plate stock : #20
    • From frozen stock : #2-3-17-5
  • Miniprep( #36-3-14-5-2-3-17-5)
  • Ligation(#20_SP-#3-14-5-2-3-17-5_XP, #1_SP-#3-14-5-2-3-17-5_XP)
  • Direct Ligation
    • i.PCR amplification -> Digest for >1h -> Heat kill for 20min (#20-3-37-5_XP, #36-3-14-5-2-3-17-5_ES)
    • ii.Digest for >1h -> Heat kill for 20min (#36-3-14-5-2-3-17-5_SP, pSB1A3_EP, #2-3-17-5_EX)
    • iii.Ligation for 30min (#20-3-14-5_ES-#2-3-17-5_EX, #36-3-14-5-2-3-17-5_SP-#20-3-37-5_XP, #36-3-14-5-2-3-17-5_ES-#20-3-37-5_XP-pSB1A3_EP)
    • iv.Transformation
  • Making 0.125% agar LB amp plate (IPTG 1, 10, 30, 100uM)
  • Cell Diffusion Assay
    • #20-3-29-5 WT (IPTG 1, 10, 30, 100uM)
    • #20-3-29-5 cheZ-/- (IPTG 1, 10, 30, 100uM)
    • WT
    • cheZ-/-

'11/9/12 (Mon)

  • Colony PCR
    • #20-#3-14-5-2-3-17-5
    • #20-3-14-5-#2-3-17-5
    • #1-#3-14-5-2-3-17-5
    • #36-3-14-5-2-3-17-5-#20-3-37-5 (no colony)
    • #36-3-14-5-2-3-17-5-#20-3-37-5-pSB1A3 (PCR failed?)

Note: #20-3-37-5 colonies on the master-plate are RED. Maybe #2-3-11-5...
Note: #36-3-14-5-2-3-17-5-#20-3-37-5-pSB1A3 PCR amplification doesn't work. Seems failed extension...

  • PCR amplification
    • From master plate : #20-3-37-5
    • From 09/11 Miniprep product : #36-3-14-5-2-3-17-5

Note: Only one #20-3-37-5 colony is NOT RED.

  • Miniprep(#20, #29 (pSB1AK3), #2-3-17-5, #2-3-37-5, BBa_J04450 (pSB1C3))
  • Digest
    • From PCR : #36-3-14-5-2-3-17-5_ES, #20-3-37-5_XP
    • pSB1A3_EP, pSB1C3_EP
    • From Miniprep : #20-3-37-5_XP
    • J04450_EP, *#2-9_XP, #20-3-29-5_ES
  • Gel extraction
    • #20_SP, #36-3-14-5_SP
    • From OverNight digests : #2-3-17-5_EX
    • #36-3-14-5-2-3-17-5_ES
    • From today's PCR digests : #20-3-37-5_XP
  • Culture for Miniprep(#20-3-37-5 from 0911 master plate)
  • Dual Luciferase Assay(#20-3-14-5-2-3-17-5, #1-3-14-5-2-3-17-5, #36-3-14-5-2-3-17-5-20-3-37-5-pSB1A3)
  • Ligation
    • #36-3-14-5-2-3-17-5_ES-#20-3-37-5_XP-pSB1A3_EP
    • #36-3-14-5-2-3-17-5_ES-#20-3-37-5_XP-pSB1C3_EP
    • #3-27-5_XP-#20_SP
    • #36-3-29-5_ES-#20-3-37-5_XP-pSB1AK3_EP

'11/9/13 (Tue)

  • Colony PCR
    • #36-3-14-5-2-3-17-5_ES-#20-3-37-5_XP-pSB1A3_EP
    • #36-3-14-5-2-3-17-5_ES-#20-3-37-5_XP-pSB1C3_EP (failed)
    • #3-27-5_XP-#20_SP (failed)
    • #36-3-29-5_ES-#20-3-37-5_XP-pSB1AK3_EP
  • Culture for Luciferase Assay
    • #36-3-14-5-2-3-17-5-20-3-37-5(pSB1A3, pSB1C3) (IPTG 0, 1, 10, 38, 100uM)
    • #20-3-14-5-2-3-17-5 (IPTG 0, 1, 10, 38, 100uM)
    • #1-3-14-5-2-3-17-5 (IPTG 0uM)

Note: Seemes OD600 should NOT exceed 0.4~0.5, or intracellular luciferase will be saturated.

  • Miniprep.
    • #20-3-14-5-2-3-17-5
    • #1-3-14-5-2-3-17-5
    • #20-3-37-5 (from a non-red colony on the master plate)
  • Gel extraction
    • #20-3-37-5_XP (from non-red colony)
    • #20-3-29-5_ES(failed)
    • #2-9_XP, pSB1C3_EP

Note: #20-3-29-5 was not cut!

  • Dual Luciferase Assay
    • #1-3-14-5-2-3-17-5 (IPTG 0uM)
    • #20-3-14-5-2-3-17-5 (IPTG 0, 1, 10, 38, 100uM)
    • #36-3-14-5-2-3-17-5-20-3-37-5 (#2-3-11-5?) (IPTG 0, 1, 10, 38, 100uM)
  • Sequencing preparation
    • #20-3-37-5 (from a non-red colony on the master plate)
    • #20-3-29-5
    • #3-27-5
    • #31
  • Digest
    • #20_E
    • #3-29-5_S (from MP products) (cut check)
  • Ligation
    • #20_SP-#3-27-5_XP
  • Transformation
    • #3-14-5-2-3-17-5
    • #1-3-14-5-2-3-17-5
    • #20-3-14-5-2-3-17-5 (for plate stock)
    • #20-3-37-5 (from a non-red colony on the master plate) (for plasmid check)

'11/09/14(Wed)

  • Colony PCR
    • #3-27-5_XP-#20_SP (failed)
  • Culture for Luciferase Assay (from plate stocks)
    • #20-3-14-5-2-3-17-5 (IPTG 0, 1, 10, 38, 100uM)
    • #1-3-14-5-2-3-17-5 (IPTG 0uM)
  • Digest
    • #3-14-5, #3-17-5, #3-14-5-2-3-17-5, #1-3-14-5-2-3-17-5, #20-3-14-5-2-3-17-5 (EPcut)
    • #20_SP
  • Culture for Miniprep.
    • From plate stock : #20, #20-3-14-5-2-3-17-5, #3-14-5-2-3-17-5, #3-17-5
    • From 09/11 master plate : #3-11-5 (pSB1C3)
  • Part cloning(#19: BBa_R0010)
  • Ligation
    • #20_SP-#3-29-5_XP
    • #36-3-14-5-2-3-17-5_ES-#20-3-37-5_XP-pSB1C3
  • Making WT competent cell
  • Dual Luciferase Assay
    • #1-3-14-5-2-3-17-5 (IPTG 0uM)
    • #20-3-14-5-2-3-17-5 (IPTG 0, 1, 10, 38, 100uM)
  • Asp TLC Assay
  • Asp Diffusion Assay (0.25% Agar)
  • Fumaric Acid (Fum) Sol. preparation
  • Sequencing
    • ①F⑤R#20-3-37-5 (from a non-red colony on the master plate)
    • ②F⑥R#20-3-29-5 (#2-3-11-5?)
    • ③F⑦R#3-27-5
    • ④F⑧R#31

'11/09/15(Thu)

  • Colony PCR(#19, #36-3-14-5-2-3-17-5-20-3-37-5 (failed), #20-3-29-5)
  • Miniprep(#20, #20-3-14-5-2-3-17-5, #3-14-5-2-3-17-5, #3-17-5)
  • Gel extraction
    • #3-14-5(failed), #3-17-5, #3-14-5-2-3-17-5, #1-3-14-5-2-3-17-5, #20-3-14-5-2-3-17-5 (EPcut); #20_SP
  • Making WT competent cell
  • Making LB plates(amp:39, cm:18)
  • Culture for Miniprep(#3-11-5 (pSB1C3), #20-3-29-5, #19)
  • Ligation
    • sulAp(ES)-#3-14-5-2-3-17-5_XP-pSB1C3_PCR digest(EP)
    • recNp(ES)-#3-14-5-2-3-17-5_XP-pSB1C3_PCR digest(EP)
    • #36-3-14-5-2-3-17-5_ES-#20-3-37-5_XP-pSB1C3_PCR digest(EP)
    • #3-27-5_XP-#20_SP
    • #3-17-5_EP-pSB1C3_PCR digest(EP)
    • #3-14-5_EP-pSB1C3_PCR digest(EP)
    • #3-14-5-2-3-17-5_EP-pSB1C3_PCR digest(EP)
    • #20-3-14-5-2-3-17-5_EP-pSB1C3_PCR digest(EP)
  • Culture for Dual Luciferase Assay(#20-3-14-5-2-3-17-5)
  • TLC check (LB, M9)
  • Cell Diffusion Assay
  • Racing Assay (0.25% Agar, 10mM Asp Sol.)
  • Fumaric Acid (Fum) Sol. preparation (failed!!)
  • Sequencing

'11/09/16(Fri)

  • Colony PCR
    • sulAp(ES)-#3-14-5-2-3-17-5_XP-pSB1C3_PCR digest(EP),
    • recNp(ES)-#3-14-5-2-3-17-5_XP-pSB1C3_PCR digest(EP),
    • #36-3-14-5-2-3-17-5_ES-#20-3-37-5_XP-pSB1C3_PCR digest(EP),
    • #3-27-5_XP-#20_SP,
    • #3-17-5_EP-pSB1C3_PCR digest(EP),
    • #3-14-5_EP-pSB1C3_PCR digest(EP),
    • #3-14-5-2-3-17-5_EP-pSB1C3_PCR digest(EP),
    • #20-3-14-5-2-3-17-5_EP-pSB1C3_PCR digest(EP)
    • #36-3-14-5-2-3-17-5_ES-#20-3-37-5_XP-pSB1C3_PCR digest(EP) (retry; non-red colonies)
  • Miniprep(#3-11-5 (pSB1C3), #20-3-29-5, #19)
  • PCR amplification(#3-27-5, #36-3-14-5-2-3-17-5, pSB1C3)
  • Sequencing prep(#20-3-29-5)
  • Making agarose gel, WT competent cell,
  • Electrophoresis check(pSB1C3_PCR)
  • Digest(pSB1C3_ES, pSB1C3_EP, #3-27-5_XP, #36-3-14-5-2-3-17-5_ES)
  • Dual luciferase assay
    • #20-3-14-5-2-3-17-5 (1h, 1.5h culture)(IPTG 0, 1, 10, 100uM)(2.5mL)
  • Fumaric Acid (Fum) Sol. preparation
    • TLC test
    • Asp diffusion test
    • Asp racing assay

'11/09/17(Sat)

  • Colony PCR (retry)
    • #3-14-5, #3-17-5 (pSB1C3)
    • #36-3-14-5-2-3-17-5-20-3-37-5
  • Culture for Miniprep(#3-14-5, #3-17-5 (pSB1C3))
  • Ligation(sulAp-pSB1C3, recNp-pSB1C3, #20-#3-27-5)
  • Transformation(#20-3-29-5 into cheZ-)
  • Making medium(.25% Agar. M9& LB each 20 plates)
  • Cell diffusion assay

'11/09/18(Sun)

  • Colony PCR & Culture for Miniprep(#36-3-14-5-2-3-17-5-20-3-37-5, #33-3-14-5-2-3-17-5)
  • Miniprep(#3-14-5, #3-17-5 (pSB1C3))
  • Ligation(#20-#3-27-5, recNp-#3-14-5-2-3-17-5, sulAp-pSB1C3, recNp-pSB1C3, #31-pSB1C3)
  • Cell diffusion assay
    • Innoculate colony(WT / cheZ-/- / cheZ-/- + cheZ plasmid)
  • GFP imaging assay(Culture cells expressing GFP)

'11/09/19(Mon)

  • Culture for Dual luciferase assay(#36-3-14-5-2-3-17-5-20-3-37-5, #33-3-14-5-2-3-17-5, #1-3-14-5-2-3-17-5, #20-3-14-5-2-3-17-5)
  • Miniprep(#36-3-14-5-2-3-17-5-20-3-37-5, #33-3-14-5-2-3-17-5)
  • Ligation(#24-#3-14-5-2-3-17-5, #20-#3-27-5)
  • Sequencing(#36-3-14-5-2-3-17-5-20-3-37-5, #33-3-14-5-2-3-17-5, #20-3-29-5)
  • Dual luciferase assay(#1~, #20~, #36~, #33~ )
  • Asp racing assay(Innoculate colonies)
  • GFP imaging assay

'11/09/20(Tue)

  • colony PCR(20-3-27-5, #24-3-14-5-2-3-17-5)
  • Gel extraction(#3-27-5_XP)
  • PCR amplification -> Digest(#29_EP, #20-3-29-5_EP, #3-29-5_EP)
  • Digest(#20_SP)
  • Sequencing prep.
  • Ethanol precipitation (10ul TE)
    • #3-11-5(pSB1C3): 144.5ng/uL
    • #3-14-5: 728.0ng/uL
    • #3-17-5(pSB1C3): 505.7ng/uL
  • Ligation(#31_EP-pSB1C3_EP, #36-3-29-5_ES-#20-3-37-5_XP-pSB1C3_EP)
  • Luciferase assay
  • Asp diffusion assay

'11/09/21(Wed)

  • Making plates
  • Gel extraction(#20_SP, #29_EP, #3-29-5_EP, #20-3-29-5_EP)
  • Ligation
    • #20_SP-#3-27-5_XP, #24_SP-#3-14-5-2-3-17-5_XP, #29_EP-pSB1C3_EP, #3-29-5_EP-pSB1C3_EP, #20-3-29-5_EP-pSB1C3_EP, #3-14-5-2-3-17-5_EP-pSB1C3_EP
  • Transformation
    • #36 from BioBrick? part
    • #33-3-14-5-2-3-17-5 into BL21 comp.
  • Digest(#3-14-5-2-3-17-5_SP, #3-14-5-2-3-17-5_ES, #3-29-5_ES)
  • Luciferase Assay(#36-3-14-5-2-3-17-5)
  • Cell diffusion assay

'11/09/22(Thu)

  • Ligation (retry)
    • #3-27-5_XP-#20_SP, #3-14-5-2-3-17-5-#24_SP, #29_EP-pSB1C3_EP, #3-29-5_EP-pSB1C3_EP, #20-3-29-5_EP-pSB1C3_EP, #3-14-5-2-3-17-5_EP-pSB1C3_EP
  • Colony PCR(#36(->culture for mini prep), #33-3-14-5-2-3-17-5(->Maybe the amplification was failed))
  • Gel extraction(#3-14-5-2-3-17-5_SP, _ES; #3-29-5_ES)
  • Making plates
    • M9 0.25% Agar. +IPTG +Amp.: 20
    • LB Cm.: 20
  • Ligation
    • #3-14-5-2-3-17-5_SP-#20-3-37-5_XP,
    • #3-14-5-2-3-17-5_ES-#20-3-37-5_XP-pSB1C3,
    • #3-29-5_ES-#20-3-37-5_XP-pSB1C3_EP

'11/09/23(Fri)

  • colony PCR
  • Column purification
  • Ligation
  • PCR amplification
  • Competent cell(BL21(DE3))
  • Asp diffusion test
  • Cell diffusion assay
  • Asp chemotaxis assay

'11/09/26(Mon)

  • Ligation(#20-#3-27-5, #24-#3-14-5-2-3-17-5, recNp-3-14-5-2-3-17-5)
  • Digest(pSB1C3_ES)
  • Miniprep(#3-29-5-pSB1C3, #20-3-29-5-pSB1C3, #36)
  • PCR amplification(pSB1A3, #3-29-5)
  • Ethanol precipitation(#3-29-5-pSB1C3, #20-3-29-5-pSB1C3)

'11/09/27(Tue)

  • Colony PCR(#24-3-14-5-2-3-17-5, #20-3-27-5, #35-3-14-5-2-3-17-5)
  • Digest(#3-29-5_ES, #36_SP, pSB1C3_ES, pSB1C3_XP)
  • Ligation(#20-3-37-5_XP-#3-14-5-2-3-17-5_SP, pSB1C3_ES, #31_EP-pSB1C3_EP)
  • Gel digestion(#3-29-5_ES, #36_SP, pSB1C3_ES, pSB1C3_XP)
  • Nanodrop(#36_SP, pSB1C3)
  • PCR amplification(J04450)
  • Gel extraction(pSB1C3_ES)

'11/09/28(Wed)

  • Nanofrop
    • From PCR : #3-29-5, pSB1C3
    • From Miniprep : #3-29-5, #20-3-27-5, J04450
  • Transformation(#3-5-3-14-5-2-3-17-5) -> BL21(DE) comp.
  • PCR amplification and Digest(EPcut) (#31, #20-3-27-5)
  • Digest(pSB1C3_EP, pSB1C3_XP, #3-29-5_ES, #24_SP_SP, #24_SP)
  • Sequencing(#20-3-27-5, #3-29-5, #3-14-5, #3-17-5)
  • Making AspA culture
  • Cell diffusion assay
  • Asp taxis assay
  • Gel extraction and Nanodrop(pSB1C3_EP, pSB1C3_XP, #3-29-5_ES, #24_SP)
  • Ligation
    • #3-29-5_ES-#20-3-37-5_XP-pSB1C3_EP
    • #3-14-5-2-3-17-5_XP-pSB1C3_XP
    • #20-3-37-5_XP-pSB1C3_XP
    • #3-27-5_XP-pSB1C3_XP
    • #29_XP-pSB1C3_XP
    • #24_SP-#3-14-5-2-3-17-5_XP
    • #36_SP-#3-29-5_XP

'11/09/29(Thu)

  • Ligation(#20-3-27-5_EP-pSB1C3_EP)
  • Gel extraction and Nanodrop(#31_EP, #20-3-27-5_EP)
  • PCR amplification(pSB1C3, #3-14-5-2-3-17-5)
  • Digest(pSB1C3_EP, #3-14-5-2-3-17-5_XP)
  • Colony PCR, PCR amplification, Miniprep and Colony diffusion assay(#36_SP-#3-29-5_XP)
  • Colony PCR and Miniprep(#24-3-14-5-2-3-17-5)

'11/09/30(Fri)

  • Digest(#36-3-29-5_ES)
  • PCR amplification and Digest(EPcut)(#3-14-5-2-3-17-5, #20-3-27-5, #29, #31)
  • Digest(#3-14-5-2-3-17-5_EP, #36-3-29-5_ES, pSB1C3_EP, pSB1C3_ES)
  • Culture for MIniprep from plate stock(#3-14-5-2-3-17-5)
  • Making competent cell(JM109)
  • Gel extraction and Nanodrop(#36-3-29-5_ES_PCR, #36-3-29-5_ES_Miniprep, #3-14-5-2-3-17-5_EP, pSB1C3_EP, pSB1C3_ES, #3-14-5-2-3-17-5_EP, #20-3-27-5_EP, #29_EP, #31_EP)
  • Ligation(#3-14-5-2-3-17-5_EP-pSB1C3_EP, #20-3-27-5_EP-pSB1C3_EP, #29_EP-pSB1C3_EP, #31_EP-pSB1C3_EP, #36-3-29-5_ES-#20-3-37-5_XP-pSB1C3_EP, recNp_ES-pSB1C3_ES, sulAp_ES-pSB1C3_ES, #3-14-5-2-3-17-5_EP-pSB1C3_EP)
  • Sequencing prepartion(#20-3-27-5_F, #36-3-29-5
  • Miniprep, Digest(EXcut) and Nanodrop(#3-14-5-2-3-17-5)
  • Culture for Miniprep(pSB1C3, #36-3-29-5)
  • Making plate LB Cm

October

'11/10/01(Sat)

  • Part Sequencing for parts shipping
  • Miniprep(#36-3-29-5)
  • Sequencing preparation and Colony PCR(#20-3-27-5_F, #36-3-29-5, #24-3-14-5-2-3-17-5)
  • Colony PCR(#3-14-5-2-3-17-5)
  • second colony PCR

'11/10/02(Sun)

  • PCR amplification(using Quick Taq), Digest(Epcut), Gel extraction and Nanodrop(#3-14-5-2-3-17-5)
  • Gel extraction(#3-14-5-2-3-17-5_EX)
  • Dual Luciferase assay(#24)
  • #36-3-29-5 Assay
  • #36-3-29-5-20-3-37-5 Assay
  • Ligation(recNp_ES-#3-14-5-2-3-17-5_EX, #3-14-5-2-3-17-5_EP-pSB1C3_EP)
  • Making agarose gel
  • Asp degradation assay

'11/10/03(Mon)

  • Miniprep, Nanodrop and PCR sequencing(#20-3-27-5, #29, #31, #36-3-29-5-20-3-37-5, recNp, sulAp)
  • Colony PCR(#3-14-5-2-3-17-5-pSB1C3)
  • Preparation for shipping

'11/10/04(Tue)

  • Preparation for shipping(#3-11-5, #3-14-5, #3-17-5, #3-14-5-2-3-17-5, #20-3-27-5, #29, #3-29-5, #20-3-29-5, #36-3-29-5-20-3-37-5, #31, #33, #35)