Team:ZJU-China/Notebook/August

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Labnote

Biobrick group | Biofilm group

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July

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August

In this month we......
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September

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Week5

Day

Note

Aug.1st Monday

•Fusion PCR Vgb+YFP+tetR, NirB+RFP+tetR,

•PCR: NirB, Vgb

Aug.2nd Tuesday

•Cut: 13K+10I, 22M+10I

• Purify: 13K+10I, 22M+10I
• Ligation: Pvgb+22M+10I, Pnirb+13K+10I

• PCR backbones
• Electrophresis

• Gel excision and purification

Aug.3rd Wednesday

• Make Phusion Buffer, 5×isothermel buffer

• colony PCR: 20H, 20J, 22B

Aug.4th Thursday

• Cut: 10I+22M, 10I+13; and purification

• Ligation: Pvgb+22M+10I, Pnirb+13K+10I,
• colony PCR: 13K+10I

• Culture: 20H, 20J, 22B, 1K, 1I,3C, 5E, 7C

• Transform: 1G, 3A, 5A, 7A from the distribution plate

Aug.5th Friday

• Check the plates. Contamination, or no positive colonies

• Miniprep: 5 backbones, 22B-1, 22B-3

• PCR: nirB from 13K+10I+nirB to firm the ligation. One positive result.

• Culture the positive colony.

Aug.6th Saturday

Aug.7th Sunday

• Cut: 22M,13K with E & S

• Culture the red colonies from plate of pSB1C3

• PCR: 5 backbones
• Cut the PCR products with P+E and run the gel to confirm.

Week6

Day

Note

Aut.8th Monday

• The gel results of 5 backbones are right.

• Miniprep: pSB1C3, vgb+22M+10I

• Culture vgb+22M+1oI in hypoxia
•PCR pSB1C3

Aug.9th Tuesday

• Run the PCR product

• PCR pSB1C3 again

• Run the product, results are good.

Aug.10th Wednesday

• Culture: 20H, 20J
• Transform 13K from plate 3

• Miniprep: 20H-1, 20J-1

• Cut: the 20H-1, 20J-1, 22B-1, 22B-3 with E+P

Aug.11th Thursday

• Run the digestion results. Bands are confirmed right.

• Colony PCR vgb+22M+10I

• Cut the plasmid of vgb+22M+10I

Aug.12th Friday

• Make new stock of competent cells

• Ligation: 13K+10I
• Cut the PCR results and 22M with E+P

• Sequence: nirB, vgbL, 12I, 20H, 20J, 22B

• Transform 13K+10I+pSB1K3
• Cut 13K to validate. Results are right.

Aug.13th Saturday

• Colony PCR: 13K+10I+pSB1K3

• Culture: vgb+22M+10I in 5ml(shaking), 5ml(water bath), 10ml(water bath), 15(water bath).

• Check the YFP expression of 5ml(shaking) and 15ml(water bath). No yellow fluorescent cells,

Aug.14th Sunday

• Transform: the Gibson assembly results.

• Miniprep: 13K+10I
• Cut: 13K+10I

• Purification: the digestion results.
• Ligation: nirB+13K+10I

Week7

Day

Note

Aug.15th Monday

• Plate: nirB+13K+10I

• Colony PCR: vgb+YFP+tetR +terminater

• Gibson PCR

Aug.16th Tuesday

• Colony PCR: vgb+22M+10I, nirB+13K+10I

• No bands of 13K; 22M confirmed

Aut.17th Wednesday

• Miniprep: 22M
• Cut: 22M

• Check CFP expression. No fluorescence.

Aug.18th Thursday

• Transform: 13K, 10I, 22M, 12I, 18P

Aug.19th Friday

• Miniprep: 13K, 10I, 22M, 12I, 18P

• Cut: 13K, 10I, 22M, 12I, 18P

• Run the gel

Aug.20th Saturday

• Ligate 13K+10I, 22M+10I

Aug.21st Sunday

• Transform: the Gibson assembly results.

• Miniprep: 13K+10I
• Cut: 13K+10I

• Purification: the digestion results.
• Ligation: nirB+13K+10I

Week8

Day

Note

Aug. 22nd Monday

• Transform 13K+10I, 22M+10I

• Test new primers.
• PCR: 22M+10I, 13K+10I

• Run the PCR products.
• Cut the PCR products.

Aug. 23rd Tuesday

• Test primers: CS/CP, VF/VR, pSB1_f/r

• Cut: 11P, 1F, 22M
• Run the cut results.1F-1/2/3 are right.

• Mix the 1F-1/2 purification products

• Cut: 1F-1+1F-2(E+S), 22M-3(X+P), pSB1C3(E+P)

Aug. 24th Wednesday

Aug. 25th Thursday

Aug. 26th Friday

• Amplify: vgb, 1C3

Aug.27th Saturday

• Digest: Vgb, 1C3, fdfhF, 22M+10I, 13K+10I

Aug.28th Sunday

• Ligate: vgb+22M+10I, fdfhF+13K+10I

• Transform the ligation results.

@@ Line 51: / Line 51: @@
 <div class="main">
 <div  class="pagetitle" id="nbook">
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      <td ><h3>Labnote</h3></td>
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 <h3>Week5</h3><hr/>
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    <tr><td width="76">Day</td><td width="349">Note</td></tr>
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 Monday
 </td>
-     <td><table id="intable" width="541" border="0" cellspacing="0" cellpadding="1">
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      <td width="286">•Fusion PCR
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 </td>
-     <td><table id="intable" width="615" border="0" cellspacing="0" cellpadding="1">
+     <td><table id="intable" width="615" border="0" cellspacing="1" cellpadding="1">
        <td width="140"> •Cut: 13K+10I, 22M+10I
@@ Line 168: / Line 168: @@
 </td>
-     <td><table width="640" border="0" cellspacing="0" cellpadding="1">
+     <td><table width="640" border="0" cellspacing="1" cellpadding="1">
    <tr>
      <td width="170">• Make Phusion Buffer, 5×isothermel buffer
@@ Line 184: / Line 184: @@
 </td>
-     <td><table width="618" border="0" cellspacing="0" cellpadding="1">
+     <td><table width="618" border="0" cellspacing="1" cellpadding="1">
    <tr>
      <td >• Cut: 10I+22M, 10I+13; and purification
@@ Line 204: / Line 204: @@
 </td>
-     <td><table width="627" border="0" cellspacing="0" cellpadding="1">
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        <tr>
         <td >• Check the plates. Contamination, or no positive colonies</td>
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 </td>
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    <tr>
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 Sunday
 </td>
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        <tr>
          <td width="244">• Cut: 22M,13K with E &amp; S
@@ Line 250: / Line 250: @@
 <h3>Week6</h3><hr/>
-<table id="notesheet" width="650" border="1" cellspacing="0" cellpadding="1">
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    <tr><td width="76">Day</td><td width="349">Note</td></tr>
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 </td>
-     <td><table id="intable" width="603" border="0" cellspacing="0" cellpadding="1">
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      <td width="191">• The gel results of 5 backbones are right.
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 </td>
-     <td><table id="intable" width="615" border="0" cellspacing="0" cellpadding="1">
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        <td width="140"> • Run the PCR product
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 </td>
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      <td width="170">• Culture: 20H, 20J<br/>
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 </td>
-     <td><table width="618" border="0" cellspacing="0" cellpadding="1">
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    <tr>
      <td >• Run the digestion results. Bands are confirmed right.
@@ Line 336: / Line 336: @@
 </td>
-     <td><table width="627" border="0" cellspacing="0" cellpadding="1">
+     <td><table width="627" border="0" cellspacing="1" cellpadding="1">
        <tr>
         <td >• Make new stock of competent cells</td>
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 </td>
-     <td><table width="620" border="0" cellspacing="0" cellpadding="1">
+     <td><table width="620" border="0" cellspacing="1" cellpadding="1">
    <tr>
      <td>• Colony PCR: 13K+10I+pSB1K3</td>
@@ Line 371: / Line 371: @@
 </td>
-     <td><table width="620" border="0" cellspacing="0" cellpadding="1">
+     <td><table width="620" border="0" cellspacing="1" cellpadding="1">
        <tr>
          <td width="244">• Transform: the  Gibson assembly results.
@@ Line 392: / Line 392: @@
 <div class="block" id="nsheet">
 <h3>Week7</h3><hr/>
-<table id="notesheet" width="650" border="1" cellspacing="0" cellpadding="1">
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    <tr><td width="76">Day</td><td width="349">Note</td></tr>
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 </td>
-     <td><table id="intable" width="603" border="0" cellspacing="0" cellpadding="1">
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      <td width="191">• Plate: nirB+13K+10I
@@ Line 420: / Line 420: @@
 </td>
-     <td><table id="intable" width="615" border="0" cellspacing="0" cellpadding="1">
+     <td><table id="intable" width="615" border="0" cellspacing="1" cellpadding="1">
        <td width="140"> • Colony PCR: vgb+22M+10I, nirB+13K+10I
@@ Line 439: / Line 439: @@
 </td>
-     <td><table width="640" border="0" cellspacing="0" cellpadding="1">
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    <tr>
      <td width="170">• Miniprep: 22M<br/>
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 </td>
-     <td><table width="618" border="0" cellspacing="0" cellpadding="1">
+     <td><table width="618" border="0" cellspacing="1" cellpadding="1">
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      <td >• Transform: 13K, 10I, 22M, 12I, 18P
@@ Line 478: / Line 478: @@
 </td>
-     <td><table width="627" border="0" cellspacing="0" cellpadding="1">
+     <td><table width="627" border="0" cellspacing="1" cellpadding="1">
        <tr>
         <td >• Miniprep: 13K, 10I, 22M, 12I, 18P </td>
@@ Line 495: / Line 495: @@
 </td>
-     <td><table width="620" border="0" cellspacing="0" cellpadding="1">
+     <td><table width="620" border="0" cellspacing="1" cellpadding="1">
    <tr>
      <td>• Ligate 13K+10I, 22M+10I</td>
@@ Line 510: / Line 510: @@
 </td>
-     <td><table width="620" border="0" cellspacing="0" cellpadding="1">
+     <td><table width="620" border="0" cellspacing="1" cellpadding="1">
        <tr>
          <td width="244">• Transform: the  Gibson assembly results.
@@ Line 532: / Line 532: @@
 <h3>Week8</h3><hr/>
-<table id="notesheet" width="650" border="1" cellspacing="0" cellpadding="1">
+<table id="notesheet" width="650" border="1" cellspacing="1" cellpadding="1">
    <tr><td width="76">Day</td><td width="349">Note</td></tr>
@@ Line 542: / Line 542: @@
 </td>
-     <td><table id="intable" width="603" border="0" cellspacing="0" cellpadding="1">
+     <td><table id="intable" width="603" border="0" cellspacing="1" cellpadding="1">
      <td width="191">• Transform 13K+10I, 22M+10I
@@ Line 566: / Line 566: @@
 </td>
-     <td><table id="intable" width="615" border="0" cellspacing="0" cellpadding="1">
+     <td><table id="intable" width="615" border="0" cellspacing="1" cellpadding="1">
        <td width="140">• Test primers: CS/CP, VF/VR, pSB1_f/r
@@ Line 588: / Line 588: @@
 </td>
-     <td><table width="640" border="0" cellspacing="0" cellpadding="1">
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    <tr>
@@ Line 603: / Line 603: @@
 </td>
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    <tr>
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 </td>
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        <tr>
         <td >• Amplify: vgb, 1C3</td>
@@ Line 632: / Line 632: @@
 </td>
-     <td><table width="620" border="0" cellspacing="0" cellpadding="1">
+     <td><table width="620" border="0" cellspacing="1" cellpadding="1">
    <tr>
      <td>• Digest: Vgb, 1C3, fdfhF, 22M+10I, 13K+10I</td>
@@ Line 646: / Line 646: @@
 </td>
-     <td><table width="620" border="0" cellspacing="0" cellpadding="1">
+     <td><table width="620" border="0" cellspacing="1" cellpadding="1">
        <tr>
          <td width="244">• Ligate: vgb+22M+10I, fdfhF+13K+10I