Team:ZJU-China/Notebook/August

From 2011.igem.org

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<div class="block" id="nsheet">
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<h3>Week6</h3><hr/>
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<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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  <tr>
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    <td id="sheetleft">Aut.8th
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Monday
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</td>
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    <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>
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    <td width="249" >• Miniprep: pSB1C3, vgb+22M+10I</td><br />
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<td width="157">• Culture vgb+22M+1oI in hypoxia<br/>
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•PCR pSB1C3
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</td>
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</table></td>
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  </tr>
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  <tr>
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    <td id="sheetleft">Aug.9th
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Tuesday
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</td>
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    <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="181" > • PCR pSB1C3 again
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</td>
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  <td width="142">• Run the product, results are good.
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</td>
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</table></td>
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  </tr>
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  <tr>
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    <td id="sheetleft">Aug.10th
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Wednesday
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</td>
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    <td><table width="640" border="0" cellspacing="0" cellpadding="1">
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  <tr>
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    <td width="170">• Culture: 20H, 20J<br/>
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• Transform 13K from plate 3
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</td>
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    <td width="184">• Miniprep: 20H-1, 20J-1
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</td>
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<td>• Cut: the 20H-1, 20J-1, 22B-1, 22B-3 with E+P</td>
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  </tr>
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</table>
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</td>
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  </tr>
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  <tr>
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    <td id="sheetleft">Aug.11th
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Thursday
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</td>
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    <td><table width="618" border="0" cellspacing="0" cellpadding="1">
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  <tr>
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    <td >• Run the digestion results. Bands are confirmed right.
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</td>
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<td>• Colony PCR vgb+22M+10I
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</td>
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<td>• Cut the plasmid of vgb+22M+10I
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</td>
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  </tr>
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</table>
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</td>
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  </tr>
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  <tr>
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    <td id="sheetleft">Aug.12th
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Friday
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</td>
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    <td><table width="627" border="0" cellspacing="0" cellpadding="1">
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      <tr>
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      <td >• Make new stock of competent cells</td>
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      <td>• Ligation: 13K+10I<br/>
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• Cut the PCR results and 22M with E+P
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</td>
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      <td>• Sequence: nirB, vgbL, 12I, 20H, 20J, 22B</td>
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      <td>• Transform 13K+10I+pSB1K3<br/>
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• Cut 13K to validate. Results are right.
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</td>
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      </tr>
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    </table></td>
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  </tr>
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  <tr>
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    <td id="sheetleft">Aug.13th
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Saturday
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</td>
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    <td><table width="620" border="0" cellspacing="0" cellpadding="1">
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  <tr>
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    <td>• Colony PCR: 13K+10I+pSB1K3</td>
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    <td>• Culture: vgb+22M+10I in 5ml(shaking), 5ml(water bath), 10ml(water bath), 15(water bath).</td>
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    <td>• Check the YFP expression of 5ml(shaking) and 15ml(water bath). No yellow fluorescent cells,</td>
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  </tr>
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</table>
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</td>
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  </tr>
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  <tr>
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    <td id="sheetleft">Aug.14th
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Sunday
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</td>
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    <td><table width="620" border="0" cellspacing="0" cellpadding="1">
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      <tr>
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        <td width="244">• Transform: the  Gibson assembly results.
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</td>
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<td width="164">• Miniprep: 13K+10I<br/>
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• Cut: 13K+10I
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</td>
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<td>• Purification: the digestion results.<br/>
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• Ligation: nirB+13K+10I
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 +
</td>
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      </tr>
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    </table></td>
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  </tr>
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</table>
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<p>&nbsp;</p>
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</div>
<script type="text/javascript">
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Revision as of 05:12, 3 October 2011

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Labnote

This group...........

Week5


DayNote
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


DayNote
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