Team:ZJU-China/Notebook/August

From 2011.igem.org

(Difference between revisions)
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<p>&nbsp;</p>  
<p>&nbsp;</p>  
</div>  
</div>  
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<div class="block" id="nsheet">
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<h3>Week2</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">Jul.11th Monday</td>
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    <td><table id="intable" width="541" border="0" cellspacing="0" cellpadding="1">
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    <td width="128">•  Set up new LB culture plates with ampicillin and kanamycin</p></td>
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 +
 
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    <td width="196">•
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      ·Sterilization of Glycerol and <br />
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        ·Preparation of 25mg/mL kanamycin</td>
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  <td>•Transformation of the parts mentioned on Jul.9th for the second time</td>
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  <td>•Observation the sections</td>
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</table></td>
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  </tr>
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  <tr>
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    <td id="sheetleft">Jul 12th Tuesday
 +
</td>
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    <td><table id="intable" width="560" border="0" cellspacing="0" cellpadding="1">
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 +
    <td width="128">• Pick two colonies of each parts and cultivate them in LB medium</td>
 +
 
 +
 
 +
    <td width="203"> •Transformation of three parts(20J,20H.22B from Kit plates of 2011 Distribution ) which are related to the degradation of Cellulose </td>
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  <td width="91">•·Mini  prep to isolate 10I,12I and 22M</td>
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  <td width="130">•Conservation of 10I,12I,22M and 11P</td>
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</table></td>
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  </tr>
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  <tr>
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    <td id="sheetleft">Jul.13th
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Wednesday
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</td>
 +
    <td><table width="640" border="0" cellspacing="0" cellpadding="1">
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  <tr>
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    <td width="120">• Place the culture plate of 20J,20H and 22B in the fridge.</td>
 +
    <td width="102">•Min prep to isolate 13K
 +
•Conservation of 13K
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</td>
 +
<td width="156">•Restriction digest of the parts(12I,10I,22M,13K) with EcoRI and PstI</td>
 +
<td width="95">•Gel electrophoresis to analyse restriction fragments</td>
 +
<td width="157">•Test the Tm of Primers CP1&amp;CS,NP&amp;NS with 13K. The result of Gel electrophoresis shows that 60.2℃ is the Tm of NS and NP, and 57.4℃ is the Tm of CS and CP1.</td>
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  </tr>
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</table>
 +
</td>
 +
  </tr>
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  <tr>
 +
    <td id="sheetleft">Jul.14th
 +
Thursday
 +
</td>
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    <td><table width="618" border="0" cellspacing="0" cellpadding="1">
 +
  <tr>
 +
    <td >• Test the Tm of the YFP,RFP,tetR,Vgb and nirB primers.
 +
The result of Gel electrophoresis shows that the Tm for the primers of Vgb is 54℃ and the Tm for the primers of nirB is 55℃.The RCR of YFP,RFP and tetR failed.
 +
</td>
 +
 
 +
  </tr>
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</table>
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</td>
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  </tr>
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  <tr>
 +
    <td id="sheetleft">Jul.15th
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Friday
 +
</td>
 +
    <td><table width="600" border="0" cellspacing="0" cellpadding="1">
 +
      <tr>
 +
        <td width="155"> • PCR(Phusion)<br/>
 +
• Digest 10I, 22M, 13K
 +
• Used wrong cutter, digestion again.
 +
</td>
 +
<td>• Run the results of PCR and the first digestion. The annealing temperature of YFP needed change. The digestion results confirmed</td>
 +
<td>• Run the digestion results of second time. The bands are confirmed.</td>
 +
      </tr>
 +
    </table></td>
 +
  </tr>
 +
  <tr>
 +
    <td id="sheetleft">Jul.16th
 +
Saturday
 +
</td>
 +
    <td><table width="620" border="0" cellspacing="0" cellpadding="1">
 +
  <tr>
 +
    <td width="127">• Cut the linearized pSB1k3 with E+P</td>
 +
    <td width="123">• Purify the digestion results of 22M, 13K, 10I</td>
 +
    <td width="133">• Confirm digestion of pSB1k3 by electrophoresis, then purification</td>
 +
    <td width="113">• Test Tm of YFP<br/>
 +
• ligation: 22M+10I, 13K+10I
 +
</td>
 +
<td width="114">• Tm of YFP is 54 degree
 +
• transform the ligation results.
 +
</td>
 +
  </tr>
 +
</table>
 +
</td>
 +
  </tr>
 +
  <tr>
 +
    <td id="sheetleft">Jul.17th
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Sunday
 +
</td>
 +
    <td><table width="620" border="0" cellspacing="0" cellpadding="1">
 +
      <tr>
 +
        <td>• PCR 22M<br/>
 +
• purify Vgb, RFP, RFP-tetR, YFP-tetR. Gibson PCR.
 +
</td>
 +
<td>• ligation the purified the fragments in yesterday.</td>
 +
<td>• 22M PCR<br/>
 +
• Transform the ligation results.
 +
</td>
 +
      </tr>
 +
    </table></td>
 +
  </tr>
 +
 +
</table>
 +
<p>&nbsp;</p>
<script type="text/javascript">
<script type="text/javascript">

Revision as of 17:34, 2 October 2011

<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" "http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd">

Labnote

 Biobrick group | Biofilm group 

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

Week1

DayNote
Jul.4th Monday
• Aerobic cultivation of DH5α • Preparation of apparatus for the formation of biofilm
Jul 5th Tuesday  
Jul.6th Wednesday
•Receiving primers ordered previously
Jul.7th Thursday
• Preparation of the aliquot of the primers • Something wrong with a shaking incubator
Jul.8th Friday
• Preparation of culture plates for the transformations
Jul.9th Saturday
• Preparation of culture plates for the transformations • protocols of transformation
Jul.10th Sunday
• Several colonies were picked up and cultivated in 5mL LB medium. •Cryosectioning of biofilm

 

Week2

DayNote
Jul.11th Monday
• Set up new LB culture plates with ampicillin and kanamycin

 • ·Sterilization of Glycerol and
·Preparation of 25mg/mL kanamycin		 •Transformation of the parts mentioned on Jul.9th for the second time •Observation the sections
Jul 12th Tuesday
• Pick two colonies of each parts and cultivate them in LB medium •Transformation of three parts(20J,20H.22B from Kit plates of 2011 Distribution ) which are related to the degradation of Cellulose •·Mini prep to isolate 10I,12I and 22M •Conservation of 10I,12I,22M and 11P
Jul.13th Wednesday
• Place the culture plate of 20J,20H and 22B in the fridge. •Min prep to isolate 13K •Conservation of 13K •Restriction digest of the parts(12I,10I,22M,13K) with EcoRI and PstI •Gel electrophoresis to analyse restriction fragments •Test the Tm of Primers CP1&CS,NP&NS with 13K. The result of Gel electrophoresis shows that 60.2℃ is the Tm of NS and NP, and 57.4℃ is the Tm of CS and CP1.
Jul.14th Thursday
• Test the Tm of the YFP,RFP,tetR,Vgb and nirB primers. The result of Gel electrophoresis shows that the Tm for the primers of Vgb is 54℃ and the Tm for the primers of nirB is 55℃.The RCR of YFP,RFP and tetR failed.
Jul.15th Friday
• PCR(Phusion)
• Digest 10I, 22M, 13K • Used wrong cutter, digestion again. 		• Run the results of PCR and the first digestion. The annealing temperature of YFP needed change. The digestion results confirmed • Run the digestion results of second time. The bands are confirmed.
Jul.16th Saturday
• Cut the linearized pSB1k3 with E+P • Purify the digestion results of 22M, 13K, 10I • Confirm digestion of pSB1k3 by electrophoresis, then purification • Test Tm of YFP
• ligation: 22M+10I, 13K+10I 		• Tm of YFP is 54 degree • transform the ligation results.
Jul.17th Sunday
• PCR 22M
• purify Vgb, RFP, RFP-tetR, YFP-tetR. Gibson PCR. 		• ligation the purified the fragments in yesterday. • 22M PCR
• Transform the ligation results.

 

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