Team:UT-Tokyo/LabNote

From 2011.igem.org

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=Parts List=
=Parts List=
-
{|
+
 
-
|+ Table 1. List of iGEM parts
+
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"
 +
|+ 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
|-  
|-  
-
| #33 || - || sulAp || pSB1AK3 || class="number" 67
+
| #33 || - || sulAp || pSB1AK3 || class="number" | 67
|-  
|-  
-
| #34 || - || uvrAp || pSB1AK3 || class="number" 96
+
| #34 || - || uvrAp || pSB1AK3 || class="number" | 96
|-  
|-  
| #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 59: Line 64:
|}
|}
<!--
<!--
-
| #2-9 ||  || c.Promoter (M)-RBS-GFP-d.Ter || pSB1A2 || class="number" 911
+
| #2-9 ||  || c.Promoter (M)-RBS-GFP-d.Ter || pSB1A2 || class="number" | 911
|-  
|-  
-
| #20-9 ||  || lacP-RBS-GFP-d.Ter || pSB1A2 || class="number" 931
+
| #20-9 ||  || lacP-RBS-GFP-d.Ter || pSB1A2 || class="number" | 931
|-  
|-  
-
| #22-9 ||  || pT7-RBS-GFP-d.Ter || pSB1AK8 || class="number" 922
+
| #22-9 ||  || pT7-RBS-GFP-d.Ter || pSB1AK8 || class="number" | 922
|-  
|-  
-
| #3-11 ||  || RBS-RFP || pSB1A2 || class="number" 736
+
| #3-11 ||  || RBS-RFP || pSB1A2 || class="number" | 736
|-  
|-  
-
| #3-11-5 ||  || RBS-RFP-d.Ter || pSB1AK3 || class="number" 831
+
| #3-11-5 ||  || RBS-RFP-d.Ter || pSB1AK3 || class="number" | 831
|-  
|-  
-
|  ||  ||  || pSB1C3 || class="number" 831
+
|  ||  ||  || pSB1C3 || class="number" | 831
|-  
|-  
-
| #2-3-11-5 ||  || c.Promoter(M)-RBS-RFP-d.Ter || pSB1A2 || class="number" 866
+
| #2-3-11-5 ||  || c.Promoter(M)-RBS-RFP-d.Ter || pSB1A2 || class="number" | 866
|-  
|-  
-
|  ||  ||  || pSB1AK3 || class="number" 866
+
|  ||  ||  || pSB1AK3 || class="number" | 866
|-  
|-  
-
| #24-3-11-5 ||  || recAp-RBS-RFP-d.Ter || pSB1A2 || class="number" 1023
+
| #24-3-11-5 ||  || recAp-RBS-RFP-d.Ter || pSB1A2 || class="number" | 1023
|-  
|-  
-
| #33-3-11-5 ||  || sulAp-RBS-RFP-d.Ter || pSB1AK3 || class="number" 898
+
| #33-3-11-5 ||  || sulAp-RBS-RFP-d.Ter || pSB1AK3 || class="number" | 898
|-  
|-  
-
| #14-5 ||  || fLuc-d.Ter || pSB1AK3 || class="number" 1748
+
| #14-5 ||  || fLuc-d.Ter || pSB1AK3 || class="number" | 1748
|-  
|-  
-
| #3-14-5 ||  || RBS-fLuc-d.Ter || pSB1A2 || class="number" 1761
+
| #3-14-5 ||  || RBS-fLuc-d.Ter || pSB1A2 || class="number" | 1761
|-  
|-  
-
| #2-3-14-5 ||  || c.Promoter (M)-RBS-fLuc-d.Ter  || pSB1A2 || class="number" 1796
+
| #2-3-14-5 ||  || c.Promoter (M)-RBS-fLuc-d.Ter  || pSB1A2 || class="number" | 1796
|-  
|-  
-
| #20-3-14-5 ||  || lacP-RBS-fLuc-d.Ter || pSB1A2 || class="number" 1816
+
| #20-3-14-5 ||  || lacP-RBS-fLuc-d.Ter || pSB1A2 || class="number" | 1816
|-  
|-  
-
| #34-3-14-5 ||  || uvrAp-RBS-fLuc-d.Ter || pSB1AK3 || class="number" 1857
+
| #34-3-14-5 ||  || uvrAp-RBS-fLuc-d.Ter || pSB1AK3 || class="number" | 1857
|-  
|-  
-
| #36-3-14-5 ||  || cIp-RBS-fLuc-d.Ter || pSB1A2 || class="number" 1810
+
| #36-3-14-5 ||  || cIp-RBS-fLuc-d.Ter || pSB1A2 || class="number" | 1810
|-  
|-  
-
| #17-5 ||  || rLuc-d.Ter  || pSB1AK3 || class="number" 1031
+
| #17-5 ||  || rLuc-d.Ter  || pSB1AK3 || class="number" | 1031
|-  
|-  
-
| #3-17-5 ||  || RBS-rLuc-d.Ter  || pSB1A2 || class="number" 1044
+
| #3-17-5 ||  || RBS-rLuc-d.Ter  || pSB1A2 || class="number" | 1044
|-  
|-  
-
| #2-3-17-5 ||  || c.Promoter (M)-RBS-rLuc-d.Ter  || pSB1A2 || class="number" 1079
+
| #2-3-17-5 ||  || c.Promoter (M)-RBS-rLuc-d.Ter  || pSB1A2 || class="number" | 1079
|-  
|-  
-
| #24-3-17-5 ||  || recAp-RBS-rLuc-d.Ter || pSB1A2 || class="number" 1236
+
| #24-3-17-5 ||  || recAp-RBS-rLuc-d.Ter || pSB1A2 || class="number" | 1236
|-  
|-  
-
| #34-3-17-5 ||  || uvrAp-RBS-rLuc-d.Ter || pSB1AK3 || class="number" 1140
+
| #34-3-17-5 ||  || uvrAp-RBS-rLuc-d.Ter || pSB1AK3 || class="number" | 1140
|-  
|-  
-
| #20-3-17-5 ||  || lacP-RBS-rLuc-d.Ter  || pSB1A2 || class="number" 1099
+
| #20-3-17-5 ||  || lacP-RBS-rLuc-d.Ter  || pSB1A2 || class="number" | 1099
|-  
|-  
-
| #3-14-5-2-3-17-5 ||  || RBS-fLuc-d.Ter-c.Promoter (M)-RBS-rLuc-d.Ter  || pSB1A2 || class="number" 2840
+
| #3-14-5-2-3-17-5 ||  || RBS-fLuc-d.Ter-c.Promoter (M)-RBS-rLuc-d.Ter  || pSB1A2 || class="number" | 2840
|-  
|-  
-
| #20-3-14-5-2-3-17-5 ||  || lacP-RBS-fLuc-d.Ter-c.Promoter (M)-RBS-rLuc-d.Ter  || pSB1A2 || class="number" 2895
+
| #20-3-14-5-2-3-17-5 ||  || lacP-RBS-fLuc-d.Ter-c.Promoter (M)-RBS-rLuc-d.Ter  || pSB1A2 || class="number" | 2895
|-  
|-  
-
| #36-3-14-5-2-3-17-5 ||  || cIp-RBS-fLuc-d.Ter-c.Promoter (M)-RBS-rLuc-d.Ter  || pSB1A2 || class="number" 2889
+
| #36-3-14-5-2-3-17-5 ||  || cIp-RBS-fLuc-d.Ter-c.Promoter (M)-RBS-rLuc-d.Ter  || pSB1A2 || class="number" | 2889
|-  
|-  
-
| #3-27 ||  || RBS-AspA || pSB1A2 || class="number" 1531
+
| #3-27 ||  || RBS-AspA || pSB1A2 || class="number" | 1531
|-  
|-  
-
| #3-27-5 ||  || RBS-AspA-d.Ter || pSB1AK3 || class="number" 1626
+
| #3-27-5 ||  || RBS-AspA-d.Ter || pSB1AK3 || class="number" | 1626
|-  
|-  
-
| #20-3-27-5 ||  || lacP-RBS-AspA-d.Ter || pSB1A2 || class="number" 1681
+
| #20-3-27-5 ||  || lacP-RBS-AspA-d.Ter || pSB1A2 || class="number" | 1681
|-  
|-  
-
| #3-29 ||  || RBS-CheZ || pSB1A2 || class="number" 741
+
| #3-29 ||  || RBS-CheZ || pSB1A2 || class="number" | 741
|-  
|-  
-
| #3-29-5 ||  || RBS-CheZ-d.Ter || pSB1AK3 || class="number" 836
+
| #3-29-5 ||  || RBS-CheZ-d.Ter || pSB1AK3 || class="number" | 836
|-  
|-  
-
| #20-3-29-5 ||  || lacP-RBS-CheZ-d.Ter  || pSB1A2 || class="number" 891
+
| #20-3-29-5 ||  || lacP-RBS-CheZ-d.Ter  || pSB1A2 || class="number" | 891
|-  
|-  
-
| #36-3-29-5 ||  || cIp-RBS-CheZ-d.Ter || pSB1A2 || class="number" 885
+
| #36-3-29-5 ||  || cIp-RBS-CheZ-d.Ter || pSB1A2 || class="number" | 885
|-  
|-  
| #37-5 ||  || cI-d.Ter || pSB1AK3 || #VALUE!
| #37-5 ||  || cI-d.Ter || pSB1AK3 || #VALUE!
Line 129: Line 134:
| #20-3-37-5 ||  || lacP-RBS-cI-d.Ter || pSB1A2 || #VALUE!
| #20-3-37-5 ||  || lacP-RBS-cI-d.Ter || pSB1A2 || #VALUE!
|-  
|-  
-
| #20-28 ||  || lacP-〈T4 phage lysis device〉 || pSB1A2 || class="number" 1840
+
| #20-28 ||  || lacP-〈T4 phage lysis device〉 || pSB1A2 || class="number" | 1840
|-  
|-  
-
| #3-30 ||  || RBS-Lysis gene || pSB1A2 || class="number" 157
+
| #3-30 ||  || RBS-Lysis gene || pSB1A2 || class="number" | 157
|-  
|-  
-
| #3-23 ||  || RBS-T7 RNA Pol. || pSB1A2 || class="number" 2668
+
| #3-23 ||  || RBS-T7 RNA Pol. || pSB1A2 || class="number" | 2668
|-  
|-  
-
| pSB1A2 ||  || Plasmid backbone || pSB1A2 || class="number" 2079
+
| pSB1A2 ||  || Plasmid backbone || pSB1A2 || class="number" | 2079
|-  
|-  
-
| pSB1AK3 ||  || Plasmid backbone || pSB1AK3 || class="number" 3189
+
| pSB1AK3 ||  || Plasmid backbone || pSB1AK3 || class="number" | 3189
|-  
|-  
-
| pSB1A3 ||  || Linearized plasmid backbone || pSB1A3 || class="number" 2155
+
| pSB1A3 ||  || Linearized plasmid backbone || pSB1A3 || class="number" | 2155
|-  
|-  
-
| pSB1K3 ||  || Linearized plasmid backbone || pSB1K3 || class="number" 2204
+
| pSB1K3 ||  || Linearized plasmid backbone || pSB1K3 || class="number" | 2204
|-  
|-  
-
| pSB1C3 ||  || Linearized plasmid backbone || pSB1C3 || class="number" 2070
+
| pSB1C3 ||  || Linearized plasmid backbone || pSB1C3 || class="number" | 2070
|-  
|-  
-
| pSB1C3 || J04450 || lacP-RBS-RFP-d.Ter || pSB1C3 || class="number" 1069
+
| pSB1C3 || J04450 || lacP-RBS-RFP-d.Ter || pSB1C3 || class="number" | 1069
|}
|}
-->
-->
 +
=Lab Notebook=
 +
 +
<html>
 +
 +
<ul id="months">
 +
<li>May</li>
 +
<li>June</li>
 +
<li>July</li>
 +
<li>August</li>
 +
<li>September</li>
 +
<li>October</li>
 +
</ul><br />
 +
 +
<div id="calendar">
 +
  <p>Please enable Javascript to view this calendar.</p>
 +
</div>
 +
 +
<div id="labnote">
 +
</html>
 +
 +
<html><div class="labMonth" id="labMonth_5"></html>
 +
==May==
 +
<html><div class="labDay"></html>
 +
==='11/05/18 (Wed)===
 +
*Making LB medium, 50×TAE, Tris-HCl (pH8.0)
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/05/24 (Tue)===
 +
*Making SOB medium, 0.5M EDTA (pH8.0)
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/05/25(Wed)===
 +
*Making TB(pH=6.7), LB plate
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/05/26(Thu)===
 +
*Making Mgaq(for SOB medium), Competent cell
 +
*TB filtration
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/05/31(Tue)===
 +
*iGEM parts resuspension + frozen stock (at -20℃)
 +
*Transformation
 +
*Overnight culture on LB plate (with 100ug/ml ampicilline)
 +
<html></div></html>
 +
<html></div></html>
 +
 +
<html><div class="labMonth" id="labMonth_6"></html>
 +
==June==
 +
<html><div class="labDay"></html>
 +
==='11/06/01(Wed)===
 +
*Picking up colony and transfer to LB medium
 +
*Making LB medium
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/06/02(Thu)===
 +
*Miniprep
 +
*Making Glycerol(50%)(for cryopreservation)
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/06/07(Tue)===
 +
*Nanodrop
 +
*Transformation
 +
*Culture from frozen stock
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/06/08(Wed)===
 +
*Picking up colony
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/06/09(Thu)===
 +
*Miniprep
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/06/13(Mon)===
 +
*Planting Negative control
 +
*Making frozen stock
 +
*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 class="labDay"></html>
 +
==='11/06/14(Tue)===
 +
*Picking up colony
 +
*Making Mg reagent
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/06/15(Wed)===
 +
*Miniprep
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/06/17(Fri)===
 +
*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><html><span class="part_num">5</span></html>)
 +
*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 class="labDay"></html>
 +
==='11/06/21(Tue)===
 +
*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><html><span class="part_num">9</span></html>, J31000)
 +
*Nanodrop again
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/06/22(Wed)===
 +
*Miniprep(<nowiki>#</nowiki><html><span class="part_num">9</span></html>, J31000)
 +
*Making competent cell
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/06/24(Fri)===
 +
*Digest (product of Miniprep 06/09 EScut)
 +
*Making agarose gel
 +
*Electrophoresis
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/06/28(Tue)===
 +
*Digest(product of Miniprep 06/09 EPcut)
 +
*Electrophoresis
 +
*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),
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/06/29(Wed)===
 +
*Picking up colony
 +
*Making LB broth
 +
<html></div></html>
 +
<html></div></html>
 +
 +
<html><div class="labMonth" id="labMonth_7"></html>
 +
==July==
 +
<html><div class="labDay"></html>
 +
==='11/07/01(Fri)===
 +
*Digest(EPcut)
 +
*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 class="labDay"></html>
 +
==='11/07/05(Tue)===
 +
*Electrophoresis(product of digest 07/01)
 +
*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)
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/07/06(Wed)===
 +
*Gel  extraction (pre)(BBa_I712052)
 +
*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
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/07/07(Thu)===
 +
*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><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><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)
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/07/08(Fri)===
 +
*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 class="labDay"></html>
 +
==='11/07/11(Mon)===
 +
*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><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 class="labDay"></html>
 +
==='11/07/12(Tue)===
 +
*Picking up colony
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/07/13(Wed)===
 +
*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><html><span class="part_num">11</span></html>)
 +
*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><html><span class="part_num">2</span></html> SPcut)
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/07/14(Thu)===
 +
*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><html><span class="part_num">2</span></html>)
 +
*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><html><span class="part_num">11</span></html>)
 +
*Making LB+amp
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/07/15(Fri)===
 +
*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><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
 +
*Frozen stock(<nowiki>#</nowiki><html><span class="part_num">11</span></html>)
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/07/19(Tue)===
 +
*Digest
 +
*Gel extraction
 +
*Making competent cell
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/07/20(Wed)===
 +
*Picking up colony
 +
*Making Master plate
 +
*Testing product of Miniprep
 +
*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 class="labDay"></html>
 +
==='11/07/21(Thu)===
 +
*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
 +
*Digest
 +
**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><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
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/07/22(Fri)===
 +
*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><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
 +
**SPcut : <nowiki>#</nowiki><html><span class="part_num">20</span></html>
 +
**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><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 class="labDay"></html>
 +
==='11/07/25(Mon)===
 +
*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><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><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×)
 +
*Dispensing PCR Mix
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/07/26(Tue)===
 +
*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><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><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><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><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)
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/07/27(Wed)===
 +
*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
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/07/28(Thu)===
 +
*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
 +
**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><html><span class="part_num">3</span>-<span class="part_num">11</span></html>
 +
**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><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
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/07/29(Fri)===
 +
*Cloning(lexA, cheZ)
 +
*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><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><html><span class="part_num">30</span></html>)
 +
*Digest(lexA, cheZ PCR product XP cut)
 +
<html></div></html>
 +
<html></div></html>
 +
 +
<html><div class="labMonth" id="labMonth_8"></html>
 +
==August==
 +
<html><div class="labDay"></html>
 +
==='11/08/01(Mon)===
 +
*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><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)
 +
*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><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><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 class="labDay"></html>
 +
==='11/08/02(Tue)===
 +
*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><html><span class="part_num">27</span></html>, <nowiki>#</nowiki><html><span class="part_num">28</span></html>)
 +
*Gel extraction
 +
*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><html><span class="part_num">30</span></html> again)
 +
*Making competent cell(cheZ-, Wild Type)
 +
*Miniprep
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/08/04(Thu)===
 +
*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><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
 +
*Digest(<nowiki>#</nowiki><html><span class="part_num">23</span></html> XPcut)
 +
*IPTG induct(<nowiki>#</nowiki><html><span class="part_num">20</span>-<span class="part_num">9</span></html>)
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/08/05(Fri)===
 +
*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><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 class="labDay"></html>
 +
==='11/08/08(Mon)===
 +
*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
 +
*Digest
 +
**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><html><span class="part_num">6</span></html>
 +
**EXcut : <nowiki>#</nowiki><html><span class="part_num">7</span></html>
 +
**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><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 class="labDay"></html>
 +
==='11/08/09(Tue)===
 +
*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><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><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
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/08/10(Wed)===
 +
*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)
 +
*Digest
 +
**EScut : <nowiki>#</nowiki><html><span class="part_num">6</span></html>
 +
**EXcut : <nowiki>#</nowiki><html><span class="part_num">7</span></html>
 +
**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><html><span class="part_num">23</span></html>
 +
*Defrost Primer
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/08/11(Thu)===
 +
*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)
 +
*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 class="labDay"></html>
 +
==='11/08/12(Fri)===
 +
*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)
 +
*Check Electrophoresis
 +
**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><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><html><span class="part_num">5</span></html> EXcut, <nowiki>#</nowiki><html><span class="part_num">14</span></html> EScut)
 +
*Gel extraction
 +
**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><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
 +
*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><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)
 +
*Digest
 +
**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><html><span class="part_num">3</span>-<span class="part_num">23</span></html>
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/08/15(Mon)===
 +
*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
 +
**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)
 +
*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: <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><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><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 class="labDay"></html>
 +
==='11/08/16(Tue)===
 +
*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><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><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)
 +
*hotstart PCR(sulAp, uvrAp, uvrBp)
 +
*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><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><html><span class="part_num">14</span></html>, <nowiki>#</nowiki><html><span class="part_num">17</span></html>)
 +
*Culture test(<nowiki>#</nowiki><html><span class="part_num">14</span></html> on 0.3%Agar.(km))
 +
*Ethanol precipitation: 29.9ng/ul * 1000ul
 +
*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)
 +
*Waking from frozen stock(<nowiki>#</nowiki><html><span class="part_num">3</span></html>, CheZ-, WT)
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='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(<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
 +
*Gel extraction(SPcut:<nowiki>#</nowiki><html><span class="part_num">3</span></html>)
 +
*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)
 +
*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 class="labDay"></html>
 +
==='11/08/18(Thu)===
 +
*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)
 +
**From Genome :  uvrBp, cheZ
 +
**From nested-region : uvrA
 +
*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)
 +
*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><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><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><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
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/08/19(Fri)===
 +
*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><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><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
 +
*Check the list of frozen stock
 +
*Assay of recAp from biobrick parts using RFP construct & transilluminator
 +
*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 class="labDay"></html>
 +
==='11/08/22(Mon)===
 +
*Making SOB medium
 +
*Culture for Miniprep
 +
**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><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 ligation products (for mistake above)
 +
*Miniprep, Nanodrop and Electrophoresis
 +
**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><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
 +
**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><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><html><span class="part_num">33</span></html> using protocol of transformation
 +
*Culture for Frozen stock
 +
**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 class="labDay"></html>
 +
==='11/08/23(Tue)===
 +
*Gel extraction
 +
**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
 +
**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><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
 +
**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><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><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><html><span class="part_num">33</span></html> sulvage (isolation culture)
 +
*Making culture(LB amp, LB)
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/08/24(Wed)===
 +
*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><html><span class="part_num">27</span></html> (82.1 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℃
 +
**amp 32
 +
**km 11 (km conc. = 3/4 * that it was)
 +
*Miniprep and Nanodrop
 +
**<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
 +
**D-Luciferin Solution : 1ml*10 + 100ul*10 at -80℃
 +
**Coelenterazine Solution : 100ul * 10 at -20℃
 +
**Lysis buffer : 500ul * 10 at -20℃
 +
*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 class="labDay"></html>
 +
==='11/08/25(Thu)===
 +
*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><html><span class="part_num">27</span></html>, <nowiki>#</nowiki><html><span class="part_num">28</span></html> XPcut)
 +
*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><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><html><span class="part_num">14</span></html> XPcut)
 +
*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 class="labDay"></html>
 +
==='11/08/26(Fri)===
 +
*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><html><span class="part_num">33</span></html> Miniprep transformations)
 +
*Gel extraction (<nowiki>#</nowiki><html><span class="part_num">14</span></html>_XP)
 +
*Ligation (retry) (using today's competent cells, 0802 ΔcheZ cells)
 +
*Cloning (cheZ)
 +
*Digest (<nowiki>#</nowiki><html><span class="part_num">5</span></html> EXcut, cheZ PCR products XPcut)
 +
*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
 +
*Contamination check (0817 comp., 0802 comp., 0802 ⊿cheZ comp. spread on LB (amp) plates)
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='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)
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/08/28(Sun)===
 +
*Making ΔcheZ comp., LB amp plates, TB
 +
*Colony PCR & master plate preparation -> Failure!!
 +
*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><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><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><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 class="labDay"></html>
 +
==='11/08/29(Mon)===
 +
*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><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><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><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 class="labDay"></html>
 +
==='11/08/30(Tue)===
 +
*Making LB amp, Aspartic acid solution (10mM), H2O2 solution (10mM)
 +
*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
 +
*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><html><span class="part_num">27</span></html>,<nowiki>#</nowiki><html><span class="part_num">28</span></html>)
 +
*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><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><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><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><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><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><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 class="labDay"></html>
 +
==='11/08/31(Wed)===
 +
*Making LB plates(amp, km)
 +
*Luciferin reagent preparation
 +
*Flask A.C. for making competent cells
 +
*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><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><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><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><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><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
 +
**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><html><span class="part_num">36</span></html>, <nowiki>#</nowiki><html><span class="part_num">37</span></html>)
 +
**Contamination check(pSB1AK3(EPcut), pSB1K3(EPcut))
 +
**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
 +
<html></div></html>
 +
<html></div></html>
 +
 +
<html><div class="labMonth" id="labMonth_9"></html>
 +
==September==
 +
<html><div class="labDay"></html>
 +
==='11/09/01(Thu)===
 +
*Colony PCR
 +
**<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
 +
**<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)
 +
*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
 +
**<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
 +
**<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><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
 +
**<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
 +
**<nowiki>#</nowiki><html><span class="part_num">36</span></html> into Nippon Gene comp.
 +
**Ligation products into Takara comp. (including N.C.)
 +
**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><html><span class="part_num">14</span>-<span class="part_num">5</span></html> (km resistance) WT/ΔcheZ cells)
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='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.
 +
**<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><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><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.
 +
**<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><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
 +
**<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
 +
**<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><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 class="labDay"></html>
 +
==='11/09/03(Sat)===
 +
*Gel extraction
 +
*Ligation & Transformation
 +
**<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><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
 +
**Mobility assay (.5%, .25%, .125%)
 +
**Asp assay
 +
*Part Cloning (<nowiki>#</nowiki><html><span class="part_num">36</span></html>)
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/09/04(Sun)===
 +
*Hot-start PCR Pre-Mix (4×) preparation (Buffer + dNTPs + THUNDER Taq)
 +
*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
 +
*GFP Live-Imaging Assays
 +
*Ligation & Transformation (retry)
 +
**<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><html><span class="part_num">36</span></html> into Takara 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><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><html><span class="part_num">20</span></html>)
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/09/05(Mon)===
 +
*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><html><span class="part_num">20</span></html>)
 +
*Luciferase Assays
 +
**Calibrations (Firefly Luciferase standards)
 +
**Background Assay
 +
**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><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><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))
 +
*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)
 +
*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 class="labDay"></html>
 +
==='11/09/06(Tue)===
 +
*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><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><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)
 +
*Ligation & Transformation
 +
**<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><html><span class="part_num">5</span></html>, pSB1AK3 (N.C.)
 +
*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
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/09/07(Wed)===
 +
*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><html><span class="part_num">36</span></html>_SP)
 +
*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><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)
 +
*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><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><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><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
 +
**<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><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
 +
**Asp conc. check (it took 5 min at 80C)
 +
*Making M9 medium(.25, .125% Agar.)
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/09/08(Thu)===
 +
*Making competent cell (WT), LB(amp) plate
 +
*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><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><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
 +
**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><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><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><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><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><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
 +
**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
 +
*Asp Gradient test
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/09/09(Fri)===
 +
*Making competent cell (WT), TB
 +
*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
 +
**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><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
 +
**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><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><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><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
 +
**<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><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
 +
**<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><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><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))
 +
*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><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
 +
*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 class="labDay"></html>
 +
==='11/09/10(Sat)===
 +
*Culture plate storage (4℃)
 +
*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><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><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><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 class="labDay"></html>
 +
==='11/09/11(Sun)===
 +
*Digest from Miniprep products
 +
**<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>-<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
 +
**<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><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><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
 +
**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><html><span class="part_num">20</span></html>
 +
**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><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><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
 +
**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><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><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
 +
*Making 0.125% agar LB amp plate (IPTG 1, 10, 30, 100uM)
 +
*Cell Diffusion Assay
 +
**<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><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
 +
**cheZ-/-
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/9/12 (Mon)===
 +
*Colony PCR
 +
**<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><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><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><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><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><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><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
 +
**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><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><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><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
 +
**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
 +
**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><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
 +
**<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><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>_ES
 +
**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><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><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
 +
**<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><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><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">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 class="labDay"></html>
 +
==='11/9/13 (Tue)===
 +
*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>-<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><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><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><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
 +
**<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><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><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.
 +
*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>-<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">37</span>-<span class="part_num">5</span></html> (from a non-red colony on the master plate)
 +
*Gel extraction
 +
**<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><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><html><span class="part_num">2</span>-<span class="part_num">9</span></html>_XP, pSB1C3_EP
 +
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
 +
**<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><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><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
 +
**<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><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">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>
 +
**<nowiki>#</nowiki><html><span class="part_num">31</span></html>
 +
*Digest
 +
**<nowiki>#</nowiki><html><span class="part_num">20</span></html>_E
 +
**<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
 +
**<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
 +
**<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> (for plate stock)
 +
**<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 class="labDay"></html>
 +
==='11/09/14(Wed)===
 +
*Colony PCR
 +
**<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)
 +
**<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><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
 +
**<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><html><span class="part_num">20</span></html>_SP
 +
*Culture for Miniprep.
 +
**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><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><html><span class="part_num">19</span></html>: BBa_R0010)
 +
*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">29</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-pSB1C3
 +
*Making WT competent cell
 +
*Dual Luciferase Assay
 +
**<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><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 Diffusion Assay (0.25% Agar)
 +
*Fumaric Acid (Fum) Sol. preparation
 +
*Sequencing
 +
**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><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><html><span class="part_num">3</span>-<span class="part_num">27</span>-<span class="part_num">5</span></html>
 +
**④F⑧R<nowiki>#</nowiki><html><span class="part_num">31</span></html>
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/09/15(Thu)===
 +
*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><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
 +
**<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 LB plates(amp:39, cm:18)
 +
*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
 +
**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><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><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><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">17</span>-<span class="part_num">5</span></html>_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><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><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><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)
 +
*Cell Diffusion Assay
 +
*Racing Assay (0.25% Agar, 10mM Asp Sol.)
 +
*Fumaric Acid (Fum) Sol. preparation (failed!!)
 +
*Sequencing
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/09/16(Fri)===
 +
*Colony PCR
 +
**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><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><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><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">17</span>-<span class="part_num">5</span></html>_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><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><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><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><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><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><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,
 +
*Electrophoresis check(pSB1C3_PCR)
 +
*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
 +
**<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
 +
**TLC test
 +
**Asp diffusion test
 +
**Asp racing assay
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/09/17(Sat)===
 +
*Colony PCR (retry)
 +
**<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><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><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><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><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)
 +
*Cell diffusion assay
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/09/18(Sun)===
 +
*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><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><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
 +
**Innoculate colony(WT / cheZ-/- / cheZ-/- + cheZ plasmid)
 +
*GFP imaging assay(Culture cells expressing GFP)
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/09/19(Mon)===
 +
*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><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><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><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><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)
 +
*GFP imaging assay<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/09/20(Tue)===
 +
*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><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><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><html><span class="part_num">20</span></html>_SP)
 +
*Sequencing prep.
 +
*Ethanol precipitation (10ul TE)
 +
**<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><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><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><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
 +
*Asp diffusion assay
 +
<html></div></html>
 +
 +
<html><div class="labDay"></html>
 +
==='11/09/21(Wed)===
 +
*Making plates
 +
*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
 +
**<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
 +
**<nowiki>#</nowiki><html><span class="part_num">36</span></html> from BioBrick? part
 +
**<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><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><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
 +
<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 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>
 +
 +
<html></div></html>  <!-- end of div#labMonth -->
 +
 +
<html>
 +
</div> <!--  end of div#labnote -->
 +
<style>
 +
span.part_num {
 +
  cursor: pointer;
 +
  color: #fff;
 +
}
 +
 +
#calendar table {
 +
  font: 16px/20px "Segoe UI",Calibri,"Myriad Pro",Myriad,Frutiger,"Frutiger Linotype","Trebuchet MS",Helvetica,Arial,sans-serif
 +
}
 +
 +
#calendar table tr td.other-month {
 +
  color: #888;
 +
}
 +
 +
#calendar table tr td {
 +
  padding: 10px;
 +
}
 +
 +
#calendar span.clickable {
 +
  cursor: pointer;
 +
  color: #7FB9FF;
 +
}
 +
 +
ul#months li {
 +
  list-style: none;
 +
  float: left;
 +
  font-size: 18px;
 +
  color: #7FB9FF;
 +
  margin-right:20px;
 +
}
 +
 +
ul#months li.active {
 +
  font-weight: bold;
 +
  color: #fff;
 +
}
 +
 +
</style>
 +
<script type="text/javascript" src="http://igem-ut.net/2011/jquery.qtip-1.0.0-rc3.min.js"></script>
 +
<script type="text/javascript" src="http://igem-ut.net/2011/jquery.calendar-widget.js"></script>
-
=Lab Diary=
+
<script type="text/javascript">
-
For convenience sake, each part (gene, promotors, etc) are represented by consecutive  numbers (e.g. #20 for lac promotor).
+
$(function() {
-
By pointing the mouse on the part number in the labnote, you can find out the details of the part.
+
  init();
 +
});
 +
function init() {
 +
  // calendar
 +
  $('ul#months li').each(function(i) {
 +
    $(this).css('cursor','pointer').click(function() {
 +
      $('#calendar').calendarWidget({month: i+4, yaer: 2011});
 +
      $('ul#months li').removeClass('active');
 +
      $('.labMonth').hide();
 +
      $('#labMonth_'+(i+5)).show();
 +
      $('#labMonth_'+(i+5)+' .labDay h3').each(function() {
 +
        var m = $(this).text().match(/\/(\d+) *\(/);
 +
        if (!m) return;
 +
        var tag = m[1];
 +
        var target = this;
 +
        tag = tag.replace(/^0+/,'');
 +
        $('#calendar td.day'+tag+' span.day').addClass('clickable').click(function() {
 +
          $.scrollTo(target, 500);
 +
        });
 +
      });
 +
      $(this).addClass('active');
 +
    });
 +
  });
 +
  $('ul#months li').first().click();
 +
  // part description popup
 +
  var description = {};
 +
  $('#partslist tr td:first-child').each(function() {
 +
    var m = $(this).html().match(/#(\d+)/);
 +
    if (!m) return;
 +
    var num = m[1];
 +
    var msg = '';
 +
    var idx2name = { 0: 'Registry ID', 1: 'Content', 2: 'Plasmid', 3: 'Length' };
 +
    $(this).siblings().each(function(i) {
 +
      msg += idx2name[i] + ': ' + $(this).html() + '<br />';
 +
    });
 +
    description[num] = msg;
 +
  });
 +
  $('span.part_num').each(function() {
 +
    $(this).qtip({content: description[$(this).html()]});
 +
  });
 +
}
 +
</script></html>
{{:Team:UT-Tokyo/Templates/EndContent}}
{{:Team:UT-Tokyo/Templates/EndContent}}

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)