Team:ZJU-China/Notebook

From 2011.igem.org

(Difference between revisions)
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<div class="main">  
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<div id="contentwrapper">
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<<div  class="pagetitle" id="nbook">
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<div  class="pagetitle" id="nbook">
       <table style="background-color:transparent;" width="750" border="0" cellspacing="0" cellpadding="1">
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     </script>
     </script>
</div>
</div>
 +
         
 +
  <div class="bigblock" id="m_biofilm"><div class="block" id="introduction">
 +
    <h3><font color="white">Introduction</font></h3>
 +
    <hr/>
 +
    <p id="intro"><strong>abstract:</strong>.................................</p>
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<div class="bigblock"><div class="inner" id="week1">
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<div class="block" id="compartment">
-
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<h3>..........</h3>
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<div class="block" id="nsheet">  
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<hr/>
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<h3>Week1</h3><hr/>
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<p>.................
-
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</p>
-
<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.4th Monday</td>
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-
    <td><table id="intable" width="328" border="0" cellspacing="0" cellpadding="1">
+
-
 
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-
    <td width="128">• Aerobic cultivation of DH5α</td>
+
-
 
+
-
 
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    <td width="196">• Preparation of apparatus for the formation of biofilm</td>
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-
 
+
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</table></td>
+
-
  </tr>
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-
  <tr>
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    <td id="sheetleft">Jul 5th Tuesday
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-
</td>
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-
    <td>&nbsp;</td>
+
-
  </tr>
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-
  <tr>
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-
    <td id="sheetleft">Jul.6th
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-
Wednesday
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-
</td>
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    <td><table width="217" border="0" cellspacing="0" cellpadding="1">
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-
  <tr>
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    <td width="215">•Receiving  primers ordered previously</td>
+
-
   
+
-
  </tr>
+
-
</table>
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</td>
+
-
  </tr>
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-
  <tr>
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-
    <td id="sheetleft">Jul.7th
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Thursday
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-
</td>
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-
    <td><table width="238" border="0" cellspacing="0" cellpadding="1">
+
-
  <tr>
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    <td width="200">• Preparation of the aliquot of the primers</td>
+
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    <td width="200">• Something wrong with a shaking incubator</td>
+
-
  </tr>
+
-
</table>
+
-
</td>
+
-
  </tr>
+
-
  <tr>
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    <td id="sheetleft">Jul.8th
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Friday
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-
</td>
+
-
    <td><table width="222" border="0" cellspacing="0" cellpadding="1">
+
-
      <tr>
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        <td width="155">• Preparation of culture plates for the transformations</td>
+
-
      </tr>
+
-
    </table></td>
+
-
  </tr>
+
-
  <tr>
+
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    <td id="sheetleft">Jul.9th
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-
Saturday
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</td>
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    <td><table width="313" border="0" cellspacing="0" cellpadding="1">
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-
  <tr>
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    <td width="127">• Preparation of culture plates for the transformations</td>
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    <td width="182">• protocols of transformation</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">Jul.10th
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Sunday
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</td>
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    <td><table width="246" border="0" cellspacing="0" cellpadding="1">
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-
      <tr>
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        <td>• Several colonies were picked up and cultivated in  5mL LB medium.
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•Cryosectioning of biofilm
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</td>
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      </tr>
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    </table></td>
+
-
  </tr>
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-
+
-
</table>
+
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<p>&nbsp;</p>
+
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</div>
+
-
+
-
<div class="block" id="summ">
+
-
<h3 style="color:white;">Data&amp;protocol</h3>
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-
<hr/>
+
-
<div class="block" id="monday">
+
-
<h3>Monday</h3>
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-
<hr/>
+
-
<img src="https://static.igem.org/mediawiki/2011/6/61/IMG_8382.jpg" style="margin-left:100px;"  alt="blanknote" />
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<p>There is no data in first week</p>
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</div>
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<p>&nbsp;</p>
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</div>
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</div>
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<!-- 11111111111111111111111111111111111111111111111111 -->
+
-
<div class="inner" id="week2">
+
-
<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>
+
-
 
+
-
  <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">
+
-
 
+
-
    <td width="128">•  Set up new LB culture plates with ampicillin and kanamycin</p></td>
+
-
 
+
-
 
+
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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>
+
-
  <td>•Transformation of the parts mentioned on Jul.9th for the second time</td>
+
-
  <td>•Observation the sections</td>
+
-
</table></td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Jul 12th Tuesday
+
-
</td>
+
-
    <td><table id="intable" width="560" border="0" cellspacing="0" cellpadding="1">
+
-
 
+
-
    <td width="128">• Pick two colonies of each parts and cultivate them in LB medium</td>
+
-
 
+
-
 
+
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    <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>
+
-
  <td width="91">•·Mini  prep to isolate 10I,12I and 22M</td>
+
-
  <td width="130">•Conservation of 10I,12I,22M and 11P</td>
+
-
</table></td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Jul.13th
+
-
Wednesday
+
-
</td>
+
-
    <td><table width="640" border="0" cellspacing="0" cellpadding="1">
+
-
  <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
+
-
</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>
+
-
  </tr>
+
-
</table>
+
-
</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Jul.14th
+
-
Thursday
+
-
</td>
+
-
    <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>
+
-
</table>
+
-
</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Jul.15th
+
-
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
+
-
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>
+
-
+
-
+
-
</div>
+
-
<div class="block" id="summ">
+
-
<h3 style="color:white;">Data&amp;protocol</h3>
+
-
<hr/>
+
-
<div class="block" id="monday">
+
-
<h3>Jul.13th Wednesday</h3>  
+
-
<hr/>  
+
-
<p><strong>Systems of restriction digestion with EcoRI and PstI</strong></p>
+
-
+
-
<table style="margin:15px;" width="110" border="1" cellspacing="0" cellpadding="1">
+
-
  <tr>
+
-
    <td>Plasmid</td>
+
-
    <td>1μL (>100ng)</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td>EcoRI</td>
+
-
    <td>1μL</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td>PstI</td>
+
-
    <td>1μL</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td>10×Buffer Tango</td>
+
-
    <td>2μL</td>
+
-
  </tr>
+
-
  <tr>
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-
  <td>ddH2O</td>
+
-
  <td>15μL</td>
+
-
  </tr>
+
-
</table>
+
-
<img  src="https://static.igem.org/mediawiki/2011/c/c3/0713program.png"  />
+
-
+
-
<p><strong>Temperature grad</strong><br/>56℃,57.4℃,60.2℃,62.9℃,64.3℃,65.8℃</p>
+
-
+
-
<p>&nbsp;</p>
+
-
+
-
<p><strong>PCR system (test the Tm of the primers CP1&amp;CS, NP&amp;NS)</strong></p>
+
-
+
-
<table style="margin:15px;" width="110" border="1" cellspacing="0" cellpadding="1">
+
-
  <tr>
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-
    <td>10×Buffer</td>
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-
    <td> 2μL</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td>dNTPs</td>
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-
    <td>0.5μL</td>
+
-
  </tr>
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-
  <tr>
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-
    <td>primers  CP1  (NP)</td>
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-
    <td>1μL</td>
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-
  </tr>
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-
  <tr>
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-
    <td>primers  CS    (NS)</td>
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-
    <td>1μL</td>
+
-
  </tr>
+
-
  <tr>
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-
    <td>Template </td>
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-
    <td>1μL</td>
+
-
  </tr>
+
-
  <tr>
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    <td>rTaq </td>
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-
    <td>0.2μL</td>
+
-
  </tr>
+
-
  <tr>
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-
    <td>H2O </td>
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-
    <td>14.5μL</td>
+
-
  </tr>
+
-
  <tr>
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-
    <td>Total</td>
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-
    <td> 20μL</td>
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-
  </tr>
+
-
</table>
+
-
+
-
</div>
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-
<div class="block" id="tuesday">
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-
<h3>Jul.14th Thursday</h3>
+
-
<hr/>
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<p>PCR systems(test the Tm of the primers for YFP,RFP,tetR,Vgb and nirB )</p>
+
-
<table width="110" border="1" cellspacing="0" cellpadding="1">
+
-
  <tr>
+
-
    <td>10×Buffer</td>
+
-
    <td> 2μL</td>
+
-
  </tr>
+
-
  <tr>
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-
    <td>dNTPs</td>
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-
    <td>0.5μL</td>
+
-
  </tr>
+
-
  <tr>
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-
    <td>primers F  </td>
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-
    <td>0.5μL</td>
+
-
  </tr>
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-
  <tr>
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    <td>primers R </td>
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-
    <td>0.5μL</td>
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-
  </tr>
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-
  <tr>
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    <td>Template </td>
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-
    <td>1μL</td>
+
-
  </tr>
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-
  <tr>
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    <td>Taq </td>
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-
    <td>0.2μL</td>
+
-
  </tr>
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-
  <tr>
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    <td>H2O </td>
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-
    <td>15.5μL</td>
+
-
  </tr>
+
-
  <tr>
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    <td>Total</td>
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    <td> 20μL</td>
+
-
  </tr>
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-
</table>
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-
<p>&nbsp;</p>
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-
<img  src="http://ung.igem.org/wiki/images/2/20/0714program.png"  />
+
-
<p>Temperature grad and the results:</p>
+
-
<img  src="https://static.igem.org/mediawiki/2011/6/6b/0714tempera.png"  />
+
-
<p>vgb&amp;nirB pcr test:</p>
+
-
<img  src="https://static.igem.org/mediawiki/2011/7/74/20110714_vgb%26nirB_pcr_test.jpg"  />
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-
<p>&nbsp;</p>
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</div>
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<p>&nbsp;</p>
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</div>
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</div>
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-
<!-- 2222222222222222222222222222222222222222222 -->
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-
<div class="inner" id="week3">
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-
<div class="block" id="nsheet">
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-
<h3>Week3</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.18th Monday</td>
+
-
    <td><table id="intable" width="541" border="0" cellspacing="0" cellpadding="1">
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-
 
+
-
    <td width="128">• Genome extraction  of E.coli </td>
+
-
 
+
-
 
+
-
    <td width="196">•  PCR of NirB<br />
+
-
        •  cut 10I with X+P, cut 22M, 13K with E+S, cut pSB1k3 with E+P</td>
+
-
  <td>• Ligate: 10I+13K, 10I+22M <br/>
+
-
• Repeat NirB PCR
+
-
</td>
+
-
  <td><p align="left">• Culture 11P<br />
+
-
    • Miniprep 10I, 22M, 13K</p></td>
+
-
</table></td>
+
-
  </tr>
+
-
  <tr>
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-
    <td id="sheetleft">Jul 19th Tuesday
+
-
</td>
+
-
    <td><table id="intable" width="615" border="0" cellspacing="0" cellpadding="1">
+
-
 
+
-
      <td width="140">• mini-prep: 10I+13K,  10I+22M</td>
+
-
 
+
-
 
+
-
    <td width="213"> •insert 10I+13K, 10I+22M into pSB1k3 </td>
+
-
  <td width="110">•transform: 5E, 3C, 7C, 1K, 1I from the distribution plate</td>
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-
  <td width="144">•transform 10I+22M, 10I+13K</td>
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-
</table></td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Jul.20th
+
-
Wednesday
+
-
</td>
+
-
    <td><table width="640" border="0" cellspacing="0" cellpadding="1">
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-
  <tr>
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-
    <td width="170">•Colonies of 22M+10I, 13K+10I, 5E, 3C, 7C, 1K, 1I confirmed</td>
+
-
    <td width="102">• Gibson PCR
+
-
</td>
+
-
<td >•Colony PCR 22M+10I, 13K+10I, 5E, 3C, 7C, 1K, 1I</td>
+
-
<td width="95">•PCR NirB</td>
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-
  </tr>
+
-
</table>
+
-
</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Jul.21th
+
-
Thursday
+
-
</td>
+
-
    <td><table width="618" border="0" cellspacing="0" cellpadding="1">
+
-
  <tr>
+
-
    <td >•PCR verification of 22M+10I, 13K+10I, 5E, 3C, 7C, 1K, 1I
+
-
</td>
+
-
<td>• Gibson PCR
+
-
• Run the results of PCR verification. Bands confirmed.
+
-
</td>
+
-
  <td>• Culture the 22M+10I, 13K+10I, 5E, 3C, 7C, 1K, 1I
+
-
• Purification of Gibson PCR results
+
-
</td>
+
-
  </tr>
+
-
</table>
+
-
</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Jul.22th
+
-
Friday
+
-
</td>
+
-
    <td><table width="627" border="0" cellspacing="0" cellpadding="1">
+
-
      <tr>
+
-
        <td width="117"> • PCR amplification of Gibson assembly results
+
-
</td>
+
-
<td width="144">• Mini prep: 22M+10I, 22.Presever in -20<br/>
+
-
• Medi prep: 1K,1I,3C,5E,7C<br/>
+
-
• Gibson Assembly fail.
+
-
</td>
+
-
<td width="164">• PCR NirB by Phusion<br/>
+
-
•Repeat PCR by changing Pnibr to Gnirbr<br/>
+
-
•Cut the mini and medi prep results with E
+
-
</td>
+
-
<td width="194">•Run the results of PCR and digestion. Fail in PCR of NirB, succeed in 10I+3K and 10I+22M ligation.</td>
+
-
      </tr>
+
-
    </table></td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Jul.23th
+
-
Saturday
+
-
</td>
+
-
    <td><table width="620" border="0" cellspacing="0" cellpadding="1">
+
-
  <tr>
+
-
    <td >•Double digestion of 13K+10I, 22M+10I, pSB1c3
+
-
+
-
</td>
+
-
    <td >• Fail in purification of Medi prep</td>
+
-
    <td >• PCR NirB<br/>
+
-
• Ligate NirB+13K+10I, Vgb+22M+10I
+
-
</td>
+
-
+
-
  </tr>
+
-
</table>
+
-
</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Jul.24th
+
-
Sunday
+
-
</td>
+
-
    <td><table width="620" border="0" cellspacing="0" cellpadding="1">
+
-
      <tr>
+
-
        <td width="244">• Gibson assembly: NirB+RFP+tetR<br/>
+
-
• Cut results of Gibson assembly and pSB1c3.
+
-
+
-
</td>
+
-
<td width="164">• Purify the ligation results in yesterday</td>
+
-
<td width="206">• ligate with backbone
+
-
• Culture 1I, 1K, 3C, 5E, 7C, 11P
+
-
+
-
</td>
+
-
      </tr>
+
-
    </table></td>
+
-
  </tr>
+
-
+
-
</table>
+
-
<p>&nbsp;</p>
+
-
</div>
+
-
<div class="block" id="summ">
+
-
<h3 style="color:white;">Data&amp;protocol</h3>
+
-
<hr/>
+
-
<div class="block" id="monday">
+
-
<h3>Monday</h3>
+
-
<hr/>
+
-
<p>WEEK3</p>
+
-
</div>
+
-
<div class="block" id="tuesday">
+
-
<h3>Tuesday</h3>
+
-
<hr/>
+
-
<p>bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb</p></div>
+
-
<p>&nbsp;</p>
+
-
</div>
+
-
</div>
+
-
<!-- 3333333333333333333333333333333333333 -->
+
-
<div class="inner" id="week4">
+
-
<div class="block" id="nsheet">
+
-
<h3>Week4</h3><hr/>
+
-
+
-
<table id="notesheet" width="650" border="1" cellspacing="0" cellpadding="1">
+
-
  <tr><td width="76">Day</td><td width="349">Note</td></tr>
+
-
 
+
-
  <tr>
+
-
    <td id="sheetleft">Jul.25th Monday</td>
+
-
    <td><table id="intable" width="541" border="0" cellspacing="0" cellpadding="1">
+
-
 
+
-
    <td width="128">• Transform Pnirb+13k+10I+pSBK3-2</td>
+
-
 
+
-
 
+
-
    <td >• Place the culture in 4 degree</td>
+
-
  <td>• Medi-and mini-prep of five cultre
+
-
</td>
+
-
  <td>•Gibson Assembly confirmation by gel</td>
+
-
</table></td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Jul. 26th Tuesday
+
-
</td>
+
-
    <td><table id="intable" width="615" border="0" cellspacing="0" cellpadding="1">
+
-
 
+
-
      <td width="140">• Cut Pnirb, Pvgb,(E+S)<br/>
+
-
• Cut 10I+13K, 10I+22M.(X+P)
+
-
</td>
+
-
 
+
-
 
+
-
    <td width="181" > • Ligation: vgb+10I+22M, nirB+10I+13K,</td>
+
-
  <td width="142">• Ligation: pSBK13+vgb+10I+22M, pSBK13+nirB+10I+13K</td>
+
-
  <td width="144">• Transform the ligation results.</td>
+
-
</table></td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Jul.27th
+
-
Wednesday
+
-
</td>
+
-
    <td><table width="640" border="0" cellspacing="0" cellpadding="1">
+
-
  <tr>
+
-
    <td width="170">• No colony on the plate<br/>
+
-
• Gibson assembly
+
-
</td>
+
-
    <td width="184">• Run the results of Gibson assembly
+
-
</td>
+
-
<td width="280" >• Excise the bands, the purify the DNA<br/>
+
-
• Gibson Assenmly
+
-
</td>
+
-
+
-
  </tr>
+
-
</table>
+
-
</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Jul.28th
+
-
Thursday
+
-
</td>
+
-
    <td><table width="618" border="0" cellspacing="0" cellpadding="1">
+
-
  <tr>
+
-
    <td >•2011 China Meet-up    @ Hefei
+
-
</td>
+
-
+
-
  </tr>
+
-
</table>
+
-
</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Jul.29th
+
-
Friday
+
-
</td>
+
-
    <td><table width="627" border="0" cellspacing="0" cellpadding="1">
+
-
      <tr>
+
-
      <td >•2011 China Meet-up    @ Hefei</td>
+
-
      </tr>
+
-
    </table></td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Jul.30th
+
-
Saturday
+
-
</td>
+
-
    <td><table width="620" border="0" cellspacing="0" cellpadding="1">
+
-
  <tr>
+
-
   
+
-
+
-
  </tr>
+
-
</table>
+
-
</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Jul.31th
+
-
Sunday
+
-
</td>
+
-
    <td><table width="620" border="0" cellspacing="0" cellpadding="1">
+
-
      <tr>
+
-
        <td width="244">• Culture 22M+10I, 13K+10I
+
-
+
-
</td>
+
-
<td width="164">• Cut a0H, 20J, 22B</td>
+
-
+
-
      </tr>
+
-
    </table></td>
+
-
  </tr>
+
-
+
-
</table>
+
-
<p>&nbsp;</p>
+
-
</div>
+
-
+
-
<div class="block" id="summ">
+
-
+
-
<h3 style="color:white;">Data&amp;protocol</h3>
+
-
<hr/>
+
-
<div class="block" id="monday">
+
-
<h3>Monday</h3>
+
-
<hr/>
+
-
<p>WEEK3</p>
+
-
</div>
+
-
<div class="block" id="tuesday">
+
-
<h3>Tuesday</h3>
+
-
<hr/>
+
-
<p>bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb</p></div>
+
-
<p>&nbsp;</p>
+
-
</div>
+
-
</div>
+
-
<!-- 444444444444444444444444444444444 -->
+
-
<div class="inner" id="week5">
+
-
+
-
<div class="block" id="nsheet">
+
-
<h3>Week5</h3><hr/>
+
-
+
-
<table id="notesheet" width="650" border="1" cellspacing="0" cellpadding="1">
+
-
  <tr><td width="76">Day</td><td width="349">Note</td></tr>
+
-
 
+
-
  <tr>
+
-
    <td id="sheetleft">Aug.1st
+
-
Monday
+
-
</td>
+
-
    <td><table id="intable" width="541" border="0" cellspacing="0" cellpadding="1">
+
-
 
+
-
    <td width="286">•Fusion PCR
+
-
Vgb+YFP+tetR, NirB+RFP+tetR,
+
-
</td>
+
-
 
+
-
 
+
-
    <td width="251" >•PCR: NirB, Vgb</td>
+
-
 
+
-
</table></td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Aug.2nd
+
-
Tuesday
+
-
+
-
</td>
+
-
    <td><table id="intable" width="615" border="0" cellspacing="0" cellpadding="1">
+
-
 
+
-
      <td width="140"> •Cut: 13K+10I, 22M+10I
+
-
</td>
+
-
 
+
-
 
+
-
    <td width="181" > • Purify: 13K+10I, 22M+10I<br/>
+
-
• Ligation: Pvgb+22M+10I, Pnirb+13K+10I
+
-
</td>
+
-
  <td width="142">• PCR backbones<br/>
+
-
• Electrophresis
+
-
</td>
+
-
  <td width="144">• Gel excision and purification</td>
+
-
</table></td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Aug.3rd
+
-
Wednesday
+
-
+
-
</td>
+
-
    <td><table width="640" border="0" cellspacing="0" cellpadding="1">
+
-
  <tr>
+
-
    <td width="170">• Make Phusion Buffer, 5×isothermel buffer
+
-
</td>
+
-
    <td width="184">• colony PCR: 20H, 20J, 22B
+
-
</td>
+
-
+
-
  </tr>
+
-
</table>
+
-
</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Aug.4th
+
-
Thursday
+
-
+
-
</td>
+
-
    <td><table width="618" border="0" cellspacing="0" cellpadding="1">
+
-
  <tr>
+
-
    <td >• Cut: 10I+22M, 10I+13; and purification
+
-
</td>
+
-
<td>• Ligation: Pvgb+22M+10I, Pnirb+13K+10I,<br/>
+
-
• colony PCR: 13K+10I
+
-
</td>
+
-
<td>• Culture: 20H, 20J, 22B, 1K, 1I,3C, 5E, 7C</td>
+
-
+
-
<td>• Transform: 1G, 3A, 5A, 7A from the distribution plate</td>
+
-
+
-
  </tr>
+
-
</table>
+
-
</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Aug.5th
+
-
Friday
+
-
+
-
</td>
+
-
    <td><table width="627" border="0" cellspacing="0" cellpadding="1">
+
-
      <tr>
+
-
      <td >• Check the plates. Contamination, or no positive colonies</td>
+
-
      <td>• Miniprep: 5 backbones, 22B-1, 22B-3</td>
+
-
      <td>• PCR: nirB from 13K+10I+nirB to firm the ligation. One positive result.</td>
+
-
      <td>• Culture the positive colony.</td>
+
-
      </tr>
+
-
    </table></td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Aug.6th
+
-
Saturday
+
-
+
-
</td>
+
-
    <td><table width="620" border="0" cellspacing="0" cellpadding="1">
+
-
  <tr>
+
-
   
+
-
+
-
  </tr>
+
-
</table>
+
-
</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Aug.7th
+
-
Sunday
+
-
</td>
+
-
    <td><table width="620" border="0" cellspacing="0" cellpadding="1">
+
-
      <tr>
+
-
        <td width="244">• Cut: 22M,13K with E &amp; S
+
-
+
-
</td>
+
-
<td width="164">• Culture the red colonies from plate of pSB1C3</td>
+
-
<td>• PCR: 5 backbones<br/>
+
-
• Cut the PCR products with P+E and run the gel to confirm.
+
-
</td>
+
-
      </tr>
+
-
    </table></td>
+
-
  </tr>
+
-
+
-
</table>
+
-
+
-
</div>
+
-
+
-
<div class="block" id="summ">
+
-
<h3 style="color:white;">Data&amp;protocol</h3>
+
-
<hr/>
+
-
<div class="block" id="monday">
+
-
<h3>Monday</h3>
+
-
<hr/>
+
-
<p>WEEK3</p>
+
-
</div>
+
-
<div class="block" id="tuesday">
+
-
<h3>Tuesday</h3>
+
-
<hr/>
+
-
<p>bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb</p></div>
+
-
<p>&nbsp;</p>
+
-
</div>
+
-
</div>
+
-
<!-- 55555555555555555555555555555555555555 -->
+
-
<div class="inner" id="week6">
+
-
<div class="block" id="nsheet">
+
-
<h3>Week6</h3><hr/>
+
-
+
-
<table id="notesheet" width="650" border="1" cellspacing="0" cellpadding="1">
+
-
  <tr><td width="76">Day</td><td width="349">Note</td></tr>
+
-
 
+
-
  <tr>
+
-
    <td id="sheetleft">Aut.8th
+
-
Monday
+
-
+
-
</td>
+
-
    <td><table id="intable" width="603" border="0" cellspacing="0" cellpadding="1">
+
-
 
+
-
    <td width="191">• The gel results of 5 backbones are right.
+
-
</td>
+
-
 
+
-
 
+
-
    <td width="249" >• Miniprep: pSB1C3, vgb+22M+10I</td><br />
+
-
<td width="157">• Culture vgb+22M+1oI in hypoxia<br/>
+
-
•PCR pSB1C3
+
-
</td>
+
-
 
+
-
</table></td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Aug.9th
+
-
Tuesday
+
-
+
-
+
-
</td>
+
-
    <td><table id="intable" width="615" border="0" cellspacing="0" cellpadding="1">
+
-
 
+
-
      <td width="140"> • Run the PCR product
+
-
</td>
+
-
 
+
-
 
+
-
    <td width="181" > • PCR pSB1C3 again
+
-
</td>
+
-
  <td width="142">• Run the product, results are good.
+
-
</td>
+
-
+
-
</table></td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Aug.10th
+
-
Wednesday
+
-
+
-
+
-
</td>
+
-
    <td><table width="640" border="0" cellspacing="0" cellpadding="1">
+
-
  <tr>
+
-
    <td width="170">• Culture: 20H, 20J<br/>
+
-
• Transform 13K from plate 3
+
-
+
-
</td>
+
-
    <td width="184">• Miniprep: 20H-1, 20J-1
+
-
</td>
+
-
<td>• Cut: the 20H-1, 20J-1, 22B-1, 22B-3 with E+P</td>
+
-
  </tr>
+
-
</table>
+
-
</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Aug.11th
+
-
Thursday
+
-
+
-
+
-
</td>
+
-
    <td><table width="618" border="0" cellspacing="0" cellpadding="1">
+
-
  <tr>
+
-
    <td >• Run the digestion results. Bands are confirmed right.
+
-
</td>
+
-
<td>• Colony PCR vgb+22M+10I
+
-
</td>
+
-
<td>• Cut the plasmid of vgb+22M+10I
+
-
+
-
</td>
+
-
+
-
+
-
  </tr>
+
-
</table>
+
-
</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Aug.12th
+
-
Friday
+
-
+
-
+
-
</td>
+
-
    <td><table width="627" border="0" cellspacing="0" cellpadding="1">
+
-
      <tr>
+
-
      <td >• Make new stock of competent cells</td>
+
-
      <td>• Ligation: 13K+10I<br/>
+
-
• Cut the PCR results and 22M with E+P
+
-
</td>
+
-
      <td>• Sequence: nirB, vgbL, 12I, 20H, 20J, 22B</td>
+
-
      <td>• Transform 13K+10I+pSB1K3<br/>
+
-
• Cut 13K to validate. Results are right.
+
-
</td>
+
-
      </tr>
+
-
    </table></td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Aug.13th
+
-
 
+
-
Saturday
+
-
+
-
+
-
</td>
+
-
    <td><table width="620" border="0" cellspacing="0" cellpadding="1">
+
-
  <tr>
+
-
    <td>• Colony PCR: 13K+10I+pSB1K3</td>
+
-
    <td>• Culture: vgb+22M+10I in 5ml(shaking), 5ml(water bath), 10ml(water bath), 15(water bath).</td>
+
-
    <td>• Check the YFP expression of 5ml(shaking) and 15ml(water bath). No yellow fluorescent cells,</td>
+
-
+
-
  </tr>
+
-
</table>
+
-
</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Aug.14th
+
-
Sunday
+
-
+
-
</td>
+
-
    <td><table width="620" border="0" cellspacing="0" cellpadding="1">
+
-
      <tr>
+
-
        <td width="244">• Transform: the  Gibson assembly results.
+
-
+
-
</td>
+
-
<td width="164">• Miniprep: 13K+10I<br/>
+
-
• Cut: 13K+10I
+
-
</td>
+
-
<td>• Purification: the digestion results.<br/>
+
-
• Ligation: nirB+13K+10I
+
-
+
-
</td>
+
-
      </tr>
+
-
    </table></td>
+
-
  </tr>
+
-
+
-
</table>
+
-
+
-
</div>
+
-
+
-
<div class="block" id="summ">
+
-
<h3 style="color:white;">Data&amp;protocol</h3>
+
-
<hr/>
+
-
<div class="block" id="monday">
+
-
<h3>Monday</h3>
+
-
<hr/>
+
-
<p>WEEK3</p>
+
-
</div>
+
-
<div class="block" id="tuesday">
+
-
<h3>Tuesday</h3>
+
-
<hr/>
+
-
<p>bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb</p></div>
+
-
<p>&nbsp;</p>
+
-
</div>
+
-
</div>
+
-
<!-- 666666666666666666666666666666666 -->
+
-
<div class="inner" id="week7">
+
-
<div class="block" id="nsheet">
+
-
<h3>Week7</h3><hr/>
+
-
<table id="notesheet" width="650" border="1" cellspacing="0" cellpadding="1">
+
-
  <tr><td width="76">Day</td><td width="349">Note</td></tr>
+
-
 
+
-
  <tr>
+
-
    <td id="sheetleft">Aug.15th
+
-
Monday
+
-
 
+
-
 
+
-
</td>
+
-
    <td><table id="intable" width="603" border="0" cellspacing="0" cellpadding="1">
+
-
 
+
-
    <td width="191">• Plate: nirB+13K+10I
+
-
</td>
+
-
 
+
-
 
+
-
    <td width="249" >• Colony PCR: vgb+YFP+tetR +terminater</td><br />
+
-
<td width="157">• Gibson PCR
+
-
</td>
+
-
 
+
-
</table></td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Aug.16th
+
-
Tuesday
+
-
 
+
-
 
+
-
 
+
-
</td>
+
-
    <td><table id="intable" width="615" border="0" cellspacing="0" cellpadding="1">
+
-
 
+
-
      <td width="140"> • Colony PCR: vgb+22M+10I, nirB+13K+10I
+
-
</td>
+
-
 
+
-
 
+
-
    <td width="181" > • No bands of 13K; 22M confirmed
+
-
</td>
+
-
+
-
+
-
</table></td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Aut.17th
+
-
Wednesday
+
-
 
+
-
 
+
-
 
+
-
</td>
+
-
    <td><table width="640" border="0" cellspacing="0" cellpadding="1">
+
-
  <tr>
+
-
    <td width="170">• Miniprep: 22M<br/>
+
-
• Cut: 22M
+
-
 
+
-
 
+
-
</td>
+
-
    <td width="184">• Check CFP expression. No fluorescence.
+
-
</td>
+
-
 
+
-
  </tr>
+
-
</table>
+
-
</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Aug.18th
+
-
Thursday
+
-
 
+
-
 
+
-
 
+
-
</td>
+
-
    <td><table width="618" border="0" cellspacing="0" cellpadding="1">
+
-
  <tr>
+
-
    <td >• Transform: 13K, 10I, 22M, 12I, 18P
+
-
</td>
+
-
 
+
-
 
+
-
 
+
-
  </tr>
+
-
</table>
+
-
</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Aug.19th
+
-
Friday
+
-
 
+
-
 
+
-
 
+
-
</td>
+
-
    <td><table width="627" border="0" cellspacing="0" cellpadding="1">
+
-
      <tr>
+
-
      <td >• Miniprep: 13K, 10I, 22M, 12I, 18P </td>
+
-
      <td>• Cut: 13K, 10I, 22M, 12I, 18P
+
-
</td>
+
-
      <td>• Run the gel</td>
+
-
   
+
-
      </tr>
+
-
    </table></td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Aug.20th
+
-
Saturday
+
-
 
+
-
 
+
-
 
+
-
</td>
+
-
    <td><table width="620" border="0" cellspacing="0" cellpadding="1">
+
-
  <tr>
+
-
    <td>• Ligate 13K+10I, 22M+10I</td>
+
-
 
+
-
 
+
-
  </tr>
+
-
</table>
+
-
</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Aug.21st
+
-
Sunday
+
-
 
+
-
 
+
-
</td>
+
-
    <td><table width="620" border="0" cellspacing="0" cellpadding="1">
+
-
      <tr>
+
-
        <td width="244">• Transform: the  Gibson assembly results.
+
-
 
+
-
</td>
+
-
<td width="164">• Miniprep: 13K+10I<br/>
+
-
• Cut: 13K+10I
+
-
</td>
+
-
<td>• Purification: the digestion results.<br/>
+
-
• Ligation: nirB+13K+10I
+
-
 
+
-
</td>
+
-
      </tr>
+
-
    </table></td>
+
-
  </tr>
+
-
 
+
-
</table>
+
</div>
</div>
-
 
+
<div class="block" id="component">
-
<div class="block" id="summ">  
+
<h3>...........</h3>
-
<h3 style="color:white;">Data&amp;protocol</h3>  
+
<hr/>
-
<hr/>  
+
<p>....................
-
<div class="block" id="monday">  
+
</p></div>
-
<h3>Monday</h3>
+
<div class="block" id="process">
-
<hr/>
+
<h3>..............</h3>
-
<p>WEEK7</p>
+
<hr/>
-
</div>  
+
<p>...................</p></div>
-
<div class="block" id="tuesday">  
+
<p style ="clear:both"></p>
-
<h3>Tuesday</h3>  
+
-
<hr/>  
+
-
<p>bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb</p></div>
+
-
<p>&nbsp;</p>
+
-
</div>
+
-
</div>
+
-
<!-- 7777777777777777777777777777 -->
+
-
<div class="inner" id="week8">
+
-
<div class="block" id="nsheet">
+
-
<h3>Week8</h3><hr/>
+
-
 
+
-
<table id="notesheet" width="650" border="1" cellspacing="0" cellpadding="1">
+
-
  <tr><td width="76">Day</td><td width="349">Note</td></tr>
+
-
 
+
-
  <tr>
+
-
    <td id="sheetleft">Aug. 22nd
+
-
Monday
+
-
 
+
-
 
+
-
 
+
-
</td>
+
-
    <td><table id="intable" width="603" border="0" cellspacing="0" cellpadding="1">
+
-
 
+
-
    <td width="191">• Transform 13K+10I, 22M+10I
+
-
</td>
+
-
 
+
-
 
+
-
    <td width="249" >• Test new primers.<br />
+
-
• PCR: 22M+10I, 13K+10I
+
-
</td>
+
-
<td width="157">• Run the PCR products.<br />
+
-
• Cut the PCR products.
+
-
 
+
-
</td>
+
-
 
+
-
</table></td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Aug. 23rd
+
-
Tuesday
+
-
 
+
-
 
+
-
 
+
-
 
+
-
</td>
+
-
    <td><table id="intable" width="615" border="0" cellspacing="0" cellpadding="1">
+
-
 
+
-
      <td width="140">• Test primers: CS/CP, VF/VR, pSB1_f/r
+
-
</td>
+
-
 
+
-
 
+
-
    <td width="181" > • Cut: 11P, 1F, 22M<br/>
+
-
• Run the cut results.1F-1/2/3 are right.
+
-
</td>
+
-
<td>• Mix the 1F-1/2 purification products</td>
+
-
<td>• Cut: 1F-1+1F-2(E+S), 22M-3(X+P), pSB1C3(E+P)</td>
+
-
 
+
-
</table></td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Aug. 24th
+
-
Wednesday
+
-
 
+
-
 
+
-
 
+
-
 
+
-
</td>
+
-
    <td><table width="640" border="0" cellspacing="0" cellpadding="1">
+
-
  <tr>
+
-
 
+
-
  </tr>
+
-
</table>
+
-
</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Aug. 25th
+
-
Thursday
+
-
 
+
-
 
+
-
 
+
-
 
+
-
</td>
+
-
    <td><table width="618" border="0" cellspacing="0" cellpadding="1">
+
-
  <tr>
+
-
 
+
-
 
+
-
 
+
-
 
+
-
  </tr>
+
-
</table>
+
-
</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Aug. 26th
+
-
Friday
+
-
 
+
-
 
+
-
</td>
+
-
    <td><table width="627" border="0" cellspacing="0" cellpadding="1">
+
-
      <tr>
+
-
      <td >• Amplify: vgb, 1C3</td>
+
-
     
+
-
      </tr>
+
-
    </table></td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Aug.27th
+
-
Saturday
+
-
 
+
-
 
+
-
</td>
+
-
    <td><table width="620" border="0" cellspacing="0" cellpadding="1">
+
-
  <tr>
+
-
    <td>• Digest: Vgb, 1C3, fdfhF, 22M+10I, 13K+10I</td>
+
-
 
+
-
 
+
-
  </tr>
+
-
</table>
+
-
</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Aug.28th
+
-
Sunday
+
-
 
+
-
</td>
+
-
    <td><table width="620" border="0" cellspacing="0" cellpadding="1">
+
-
      <tr>
+
-
        <td width="244">• Ligate: vgb+22M+10I, fdfhF+13K+10I
+
-
 
+
-
</td>
+
-
<td width="164">• Transform the ligation results.
+
-
</td>
+
-
 
+
-
      </tr>
+
-
    </table></td>
+
-
  </tr>
+
-
 
+
-
</table>  
+
</div>
</div>
-
<div class="block" id="summ">  
+
<div class="block" id="mmodeling">
-
<h3 style="color:white;">Data&amp;protocol</h3>  
+
<h3>.............</h3><hr/>
-
<hr/>
+
<p>.................</p>
-
<div class="block" id="monday">
+
-
<h3>Monday</h3>
+
-
<hr/>  
+
-
<p>WEEK8</p>
+
-
</div>
+
-
<div class="block" id="tuesday">
+
-
<h3>Tuesday</h3>
+
-
<hr/>
+
-
<p>bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb</p></div>
+
-
<p>&nbsp;</p>
+
-
</div>
+
-
</div>
+
-
<!-- 8888888888888888888888888888 -->
+
-
<div class="inner" id="week9">
+
-
<div class="block" id="nsheet">
+
-
<h3>Week9</h3><hr/>
+
-
 
+
-
<table id="notesheet" width="650" border="1" cellspacing="0" cellpadding="1">
+
-
  <tr><td width="76">Day</td><td width="349">Note</td></tr>
+
-
 
+
-
  <tr>
+
-
    <td id="sheetleft">Aug.29th
+
-
Monday
+
-
 
+
-
 
+
-
 
+
-
 
+
-
</td>
+
-
    <td><table id="intable" width="603" border="0" cellspacing="0" cellpadding="1">
+
-
 
+
-
    <td width="191">• Meeting. Summarize all the experiment work so far.
+
-
</td>
+
-
 
+
-
 
+
-
    <td width="249" >• Cut: 22M(X+P), 1C3(E+P)
+
-
</td>
+
-
 
+
-
 
+
-
</table></td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Aug.30th
+
-
Tuesday
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
</td>
+
-
    <td><table id="intable" width="615" border="0" cellspacing="0" cellpadding="1">
+
-
 
+
-
 
+
-
 
+
-
</table></td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Aug.31st
+
-
Wednesday
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
</td>
+
-
    <td><table width="640" border="0" cellspacing="0" cellpadding="1">
+
-
  <tr>
+
-
  <td>• Cut vgb+YFP+tetR, fdhF+RFP+tetR.<br/>• Run the digestion results</td>
+
-
  <td>• Culture the positive results</td>
+
-
  </tr>
+
-
</table>
+
-
</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Sept.1st
+
-
Thursday
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
</td>
+
-
    <td><table width="618" border="0" cellspacing="0" cellpadding="1">
+
-
  <tr>
+
-
 
+
-
<td>• Miniprep: Y#4, R#2/3/4/5<br/>
+
-
• Cut Y#4, R#2/3/4/5
+
-
</td>
+
-
<td>•Run the gel.<br/>
+
-
• Send R#3,Y#4 sample to sequencing
+
-
</td>
+
-
<td>• Hypoxia culture: RFP3, RFP4<br/>
+
-
• Preserve Y#4, R#2/3/4/5
+
-
</td>
+
-
  </tr>
+
-
</table>
+
-
</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Sept.2nd
+
-
Friday
+
-
 
+
-
 
+
-
 
+
-
</td>
+
-
    <td><table width="627" border="0" cellspacing="0" cellpadding="1">
+
-
      <tr>
+
-
      <td >• Phusion PCR</td>
+
-
      <td>• Hypoxia Culture: R, Y<br/>
+
-
• Run the PCR results. Bands are confirmed right.
+
-
</td>
+
-
<td>• Purification: vgb, YFP, tetR.</td>
+
-
      </tr>
+
-
    </table></td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Sept.3rd
+
-
Saturday
+
-
 
+
-
 
+
-
 
+
-
</td>
+
-
    <td><table width="620" border="0" cellspacing="0" cellpadding="1">
+
-
  <tr>
+
-
    <td>• Culture: YFP, RFP, A25<br/>
+
-
• Culture in micro-oxygen: YFP, RFP, A25
+
-
</td>
+
-
  <td>• Culture in slope medium: RFP, YFP, A25<br/>
+
-
• Hypoxia culture: RFP, YFP, A25
+
-
</td>
+
-
<td>• Check the fluorescence.<br/>
+
-
• Neither RFP in hypoxia or oxygen condition.
+
-
Little YFP both in hypoxia and oxygen.
+
-
</td>
+
-
  </tr>
+
-
</table>
+
-
</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Sept.4th
+
-
Sunday
+
-
 
+
-
 
+
-
</td>
+
-
    <td><table width="620" border="0" cellspacing="0" cellpadding="1">
+
-
      <tr>
+
-
        <td width="244">• Miniprep: RFP3, YFP4
+
-
 
+
-
</td>
+
-
<td width="164">• Cut: RFP3, YFP4, !C3.
+
-
</td>
+
-
 
+
-
      </tr>
+
-
    </table></td>
+
-
  </tr>
+
-
 
+
-
</table>
+
</div>
</div>
-
<div class="block" id="summ">  
+
<div class="block" id="sr"></div>
-
<h3 style="color:white;">Data&amp;protocol</h3>
+
-
<hr/>
+
-
<div class="block" id="monday">
+
-
<h3>Monday</h3>
+
-
<hr/>
+
-
<p>WEEK9</p>
+
-
</div>
+
-
<div class="block" id="tuesday">
+
-
<h3>Tuesday</h3>
+
-
<hr/>
+
-
<p>bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb</p></div>
+
-
<p>&nbsp;</p>
+
-
</div>
+
-
</div>
+
-
<!-- 9999999999999999999999999999 -->
+
-
<div class="inner" id="week10">
+
-
<div class="block" id="nsheet">
+
-
<h3>Week10</h3><hr/>
+
-
<table id="notesheet" width="650" border="1" cellspacing="0" cellpadding="1">
+
-
  <tr><td width="76">Day</td><td width="349">Note</td></tr>
+
-
 
+
-
  <tr>
+
-
    <td id="sheetleft">Sept.5th
+
-
Monday
+
-
 
+
-
</td>
+
-
    <td><table id="intable" width="603" border="0" cellspacing="0" cellpadding="1">
+
-
 
+
-
    <td width="191">• Run the gel: Yu’s PCR results, 1C3, RFP3, YFP4<br/>
+
-
• Purification: RFP3, YFP4
+
-
 
+
-
</td>
+
-
 
+
-
 
+
-
    <td  >• Ligation: RFP3+1C3, YFP4+1C3<br/>
+
-
• Hypoxia culture: RFP3, YFP4, A25.
+
-
 
+
-
</td>
+
-
<td>• Transform the ligation results.<br/>
+
-
• Confirmed RFP expression only in hypoxia condition.
+
-
</td>
+
-
 
+
-
</table></td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Sept.6th
+
-
Tuesday
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
</td>
+
-
    <td><table id="intable" width="615" border="0" cellspacing="0" cellpadding="1">
+
-
  <td>• Check the RFP, YFP plates.</td>
+
-
  <td>• Colony PCR</td>
+
-
 
+
-
</table></td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Sept.7th
+
-
Wednesday
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
</td>
+
-
    <td><table width="640" border="0" cellspacing="0" cellpadding="1">
+
-
  <tr>
+
-
  <td>• Culture YFP1/2, RFP3/4</td>
+
-
 
+
-
  </tr>
+
-
</table>
+
-
</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Sept.8th
+
-
Thurday
+
-
 
+
-
 
+
-
</td>
+
-
    <td><table width="618" border="0" cellspacing="0" cellpadding="1">
+
-
  <tr>
+
-
 
+
-
<td>• Preserve the strain of RFP1, YFP1/2 in 1C3.<br/>
+
-
• Miniprep the remains.
+
-
 
+
-
</td>
+
-
<td>• Cut: YFP1/2, RFP1
+
-
</td>
+
-
<td>• Run the digestion results.<br/>
+
-
• Culture RFP2
+
-
 
+
-
</td>
+
-
  </tr>
+
-
</table>
+
-
</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Sept.9th
+
-
Friday
+
-
 
+
-
 
+
-
 
+
-
 
+
-
</td>
+
-
    <td><table width="627" border="0" cellspacing="0" cellpadding="1">
+
-
      <tr>
+
-
      <td >• Miniprep: RFP2, YFP2, RFP1</td>
+
-
      <td>• Cut them and run the gel.
+
-
</td>
+
-
 
+
-
      </tr>
+
-
    </table></td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Sept.10th
+
-
Saturday
+
-
 
+
-
 
+
-
 
+
-
 
+
-
</td>
+
-
    <td><table width="620" border="0" cellspacing="0" cellpadding="1">
+
-
  <tr>
+
-
    <td>• Cut YFP(1A3), RFP(1A3) with S+E
+
-
 
+
-
 
+
-
</td>
+
-
  <td>• Purification: RFP(1A3), YFP(1A3)
+
-
</td>
+
-
<td>• Ligation: RFP+1C3, YFP+1C3
+
-
</td>
+
-
<td>• Transform the ligation results.</td>
+
-
  </tr>
+
-
</table>
+
-
</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Sept.11th
+
-
Sunday
+
-
 
+
-
 
+
-
 
+
-
</td>
+
-
    <td><table width="620" border="0" cellspacing="0" cellpadding="1">
+
-
      <tr>
+
-
        <td width="244">• Check the plates.
+
-
 
+
-
</td>
+
-
<td width="164">• Culture lig RFY+YFP+IC3<br/>
+
-
• Culture 1K3
+
-
 
+
-
</td>
+
-
<td>• Colony PCR: RFP(1C3)， YFP(1C3)</td>
+
-
      </tr>
+
-
    </table></td>
+
-
  </tr>
+
-
 
+
-
</table>
+
-
 
+
</div>
</div>
 +
<script type="text/javascript">
 +
</script>
-
<div class="block" id="summ">
 
-
<h3 style="color:white;">Data&amp;protocol</h3>
 
-
<hr/>
 
-
<div class="block" id="monday">
 
-
<h3>Monday</h3>
 
-
<hr/>
 
-
<p>WEEK10</p>
 
-
</div>
 
-
<div class="block" id="tuesday">
 
-
<h3>Tuesday</h3>
 
-
<hr/>
 
-
<p>bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb</p></div>
 
-
<p>&nbsp;</p>
 
-
</div>
 
-
</div>
 
-
<!-- 1010101010101010101010101010 -->
 
-
<div class="inner" id="week11">
 
-
<div class="block" id="nsheet">
 
-
<h3>Week11</h3><hr/>
 
-
 
-
<table id="notesheet" width="650" border="1" cellspacing="0" cellpadding="1">
 
-
  <tr><td width="76">Day</td><td width="349">Note</td></tr>
 
    
    
-
  <tr>
 
-
    <td id="sheetleft">Sept.12th
 
-
Monday
 
-
 
-
 
-
</td>
 
-
    <td><table id="intable" width="603" border="0" cellspacing="0" cellpadding="1">
 
-
 
 
-
    <td width="191">• Colony PCR: 1A3, 1K3, lig RFP+YFP+1C3.
 
-
 
-
</td>
 
-
 
 
-
 
 
-
    <td  >• Run the PCR results.
 
-
 
-
</td>
 
-
<td>• Miniprep: lig RFP+YFP+1C3
 
-
</td>
 
-
 
 
-
</table></td>
 
-
  </tr>
 
-
  <tr>
 
-
    <td id="sheetleft">Sept.13th
 
-
Tuesday
 
-
 
-
 
-
 
-
 
-
 
-
 
-
 
-
</td>
 
-
    <td><table id="intable" width="615" border="0" cellspacing="0" cellpadding="1">
 
-
  <td>• Transform 2M, 2I, 6I, 23L.</td>
 
-
  <td>• Purification: 1K3</td>
 
-
 
-
</table></td>
 
-
  </tr>
 
-
  <tr>
 
-
    <td id="sheetleft">Sept.14th
 
-
Wednesday
 
-
 
-
 
-
 
-
 
-
 
-
 
-
 
-
</td>
 
-
    <td><table width="640" border="0" cellspacing="0" cellpadding="1">
 
-
  <tr>
 
-
  <td>• Miniprep: 20H, 20J, 22B</td>
 
-
  <td>• Cut: 1K3 with E+P, lig3 with S+E, 12I(CFP) with P+X,</td>
 
-
  <td>• Gel excision and /or purification: 1K3, lig3, 12I(CFP)</td>
 
-
  <td>• Run the gel.<br/>
 
-
• Culture: 1I, 1K, 3L, 5E, 7C
 
-
</td>
 
-
<td>• Miniprep: RFP+YFP+1C3.<br/>
 
-
• Run the gel of 1K3, lig3, CFP again.
 
-
</td>
 
-
<td>• Gel excision and purification<br/>
 
-
• Ligation: Lig3+CFP+1K3
 
-
</td>
 
-
  </tr>
 
-
</table>
 
-
</td>
 
-
  </tr>
 
-
  <tr>
 
-
    <td id="sheetleft">Sept.15th
 
-
Thursday
 
-
 
-
 
-
 
-
</td>
 
-
    <td><table width="618" border="0" cellspacing="0" cellpadding="1">
 
-
  <tr>
 
-
 
 
-
<td>• Preserve: 1I, 1K, 3C, 5E, 7C<br/>
 
-
• Transform the ligation results
 
-
</td>
 
-
<td>• Colony PCR: 2M, 2I，6I，23L<br/>
 
-
• Cut: 20H, 20J, 22B with X+P
 
-
 
-
</td>
 
-
<td>• Miniprep: 2M-1, 2I-1, 6I-1, 23L-1<br/>
 
-
• Cut: 2M, 2I with E+S
 
-
 
-
 
-
</td>
 
-
  </tr>
 
-
</table>
 
-
</td>
 
-
  </tr>
 
-
  <tr>
 
-
    <td id="sheetleft">Sept.16th
 
-
Friday
 
-
 
-
 
-
 
-
 
-
</td>
 
-
    <td><table width="627" border="0" cellspacing="0" cellpadding="1">
 
-
      <tr>
 
-
      <td >• Colony PCR: RFP+YFP+1K3</td>
 
-
      <td>• Cut 13K with E+S, 10I with P+X, 1C3 with E+P
 
-
</td>
 
-
<td>• Run the gel: 13K, 10I, 1C3, 2I, 2M, 20J, 22H, 22B</td>
 
-
<td>• Ligation: 13K+10I+1C3<br/>
 
-
• Culture: YFP(Amp), CFP(Amp)
 
-
</td>
 
-
      </tr>
 
-
    </table></td>
 
-
  </tr>
 
-
  <tr>
 
-
    <td id="sheetleft">Sept.17th
 
-
Saturday
 
-
 
-
 
-
 
-
 
-
 
-
</td>
 
-
    <td><table width="620" border="0" cellspacing="0" cellpadding="1">
 
-
  <tr>
 
-
    <td>• Order primers for 20H, 20J<br/>
 
-
• Culture 20H-1, 20J-1, 22B-1
 
-
</td>
 
-
  <td>• Plate the ligation results<br/>
 
-
• Culture RFP+YFP+CFP+1K3
 
-
 
-
</td>
 
-
<td>• Colony PCR: 3A, 1C3<br/>
 
-
• Preserve: 1K3, RFP+YFP+1C3.
 
-
 
-
</td>
 
-
  </tr>
 
-
</table>
 
-
</td>
 
-
  </tr>
 
-
  <tr>
 
-
    <td id="sheetleft">Sept.18th
 
-
Sunday
 
-
 
-
 
-
</td>
 
-
    <td><table width="620" border="0" cellspacing="0" cellpadding="1">
 
-
      <tr>
 
-
        <td width="244">• Miniprep: 20J, 20H, RFP+YFP+CFP+1K3
 
-
1A3
 
-
</td>
 
-
<td width="164">• Run the gel of PCR results
 
-
 
-
</td>
 
-
<td>• Cut RFP+YFP+CFP+1K3
 
-
with E+P<br/>
 
-
• Colony PCR: 13K+10I+1C3
 
-
</td>
 
-
<td>• PCR: fdhF+rush+Vgb+tetR<br/>
 
-
• Preserve: 20H, 20J, 22B
 
-
</td>
 
-
<td>• Run the gel: 20H, 20J, 22B<br/>
 
-
• Cut the bands and purification.
 
-
</td>
 
-
<td>• Run the gel: 13K+10I+1C3, RFP+YFP+CFP+1K3<br/>
 
-
• Culture: 13K+10I+1C3<br/>
 
-
• Grad PCR: fdhF.
 
-
</td>
 
-
      </tr>
 
-
    </table></td>
 
-
  </tr>
 
-
 
-
</table>
 
</div>
</div>
-
<div class="block" id="summ">
+
<p style ="clear:both"></p>
-
<h3 style="color:white;">Data&amp;protocol</h3>
+
<p style ="clear:both"></p>
-
<hr/>
+
           
-
<div class="block" id="monday">
+
<div class="footer" id="footer"><a href="https://2011.igem.org/"> iGem 2011 Home Page</a>
-
<h3>Monday</h3>
+
           <a href="http://ung.igem.org/Special:Upload/"> Upload Files</a>
-
<hr/>
+
           <a href="https://2011.igem.org/wiki/index.php?title=Template:Zjucss_main&action=edit">Edit CSS</a>
-
<p>WEEK11</p>
+
           <a href="http://ung.igem.org/Team_Wikis?year=2011"> Team Wikis</a>
-
</div>
+
           <a href="mailto:zjuigem@gmail.com?"><strong>Contact Us</strong></a>
-
<div class="block" id="tuesday">
+
-
<h3>Tuesday</h3>
+
-
<hr/>
+
-
<p>bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb</p></div>
+
-
<p>&nbsp;</p>
+
-
</div>
+
-
</div> 
+
-
<!-- 11 11 11 11 11 11 11 11 11  -->
+
-
<div class="inner" id="week12">
+
-
<div class="block" id="nsheet">
+
-
<h3>Week12</h3><hr/>
+
-
 
+
-
<table id="notesheet" width="650" border="1" cellspacing="0" cellpadding="1">
+
-
  <tr><td width="76">Day</td><td width="349">Note</td></tr>
+
-
 
+
-
  <tr>
+
-
    <td id="sheetleft">Sept.19th
+
-
Monday
+
-
 
+
-
 
+
-
 
+
-
</td>
+
-
    <td><table id="intable" width="603" border="0" cellspacing="0" cellpadding="1">
+
-
 
+
-
    <td width="191">• Miniprep: 13K+10I+1C3, 1C3<br/>
+
-
• Run the gel of digestion results.
+
-
 
+
-
</td>
+
-
 
+
-
 
+
-
    <td  >• Cut: 13K+10I+1C3, 1A3, fdhF
+
-
 
+
-
</td>
+
-
<td>• Run the gel: 20H, 20J, rush+vgb, RYC+1K3, fdhF, 13K+10I, 1A3
+
-
</td>
+
-
  <td>• Cut and purification: fdhF, 13K+10I, 1A3<br/>
+
-
• Culture: CFP, YFP
+
-
</td>
+
-
<td>• Purification: Rush PCR<br/>
+
-
• Cut: 22B with X, rush PCR results with S
+
-
</td>
+
-
 
+
-
</table></td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Sept.20th
+
-
Tuesday
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
</td>
+
-
    <td><table id="intable" width="615" border="0" cellspacing="0" cellpadding="1">
+
-
  <td>• Run the 20H, 20J from PCR results.<br/>
+
-
• Transform: fdhF+13K+10I+1A3
+
-
</td>
+
-
  <td>• Purification: 20B, Rush<br/>
+
-
• Ligation: 22B+Rush
+
-
</td>
+
-
<td>• Purification: ligation results<br/>
+
-
• Cut: the ligation results with X+S
+
-
</td>
+
-
<td>• PCR 20H<br/>
+
-
• Self-ligation: rush+22M
+
-
</td>
+
-
</table></td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Sept.21st
+
-
Wednesday
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
</td>
+
-
    <td><table width="640" border="0" cellspacing="0" cellpadding="1">
+
-
  <tr>
+
-
  <td>• Colony PCR: fdhF+13K+10I+1A3 on plates</td>
+
-
  <td>• Run the gel of PCR results</td>
+
-
  <td>• Culture the positive results.</td>
+
-
+
-
  </tr>
+
-
</table>
+
-
</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Sept.22nd
+
-
Thursday
+
-
 
+
-
 
+
-
 
+
-
</td>
+
-
    <td><table width="618" border="0" cellspacing="0" cellpadding="1">
+
-
  <tr>
+
-
 
+
-
<td>• Miniprep: fdhF+13K+10I+1A3<br/>
+
-
• Preserve the above.
+
-
 
+
-
</td>
+
-
<td>• Cut and run the results:
+
-
fdhF+13K+10I+1A3, 13K+10I+1C3
+
-
 
+
-
 
+
-
</td>
+
-
<td>• Cut: RFP with E+S, YFP with X+P, 1K3 with E+P, 1C3 with E+P
+
-
</td>
+
-
<td>• Purification the digestion results.</td>
+
-
  </tr>
+
-
</table>
+
-
</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Sept.23rd
+
-
Friday
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
</td>
+
-
    <td><table width="627" border="0" cellspacing="0" cellpadding="1">
+
-
      <tr>
+
-
      <td >• Ligate: RFP+1A3/ YFP+1C3/1K3, RFP+1A3/1C3</td>
+
-
      <td>• Transform the ligation results.
+
-
</td>
+
-
 
+
-
      </tr>
+
-
    </table></td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Sept. 24th
+
-
Saturday
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
</td>
+
-
    <td><table width="620" border="0" cellspacing="0" cellpadding="1">
+
-
  <tr>
+
-
    <td>• Order primers for 20H, 20J<br/>
+
-
• Culture 20H-1, 20J-1, 22B-1
+
-
</td>
+
-
  <td>• Plate the ligation results<br/>
+
-
• Culture RFP+YFP+CFP+1K3
+
-
 
+
-
</td>
+
-
<td>• Colony PCR: 3A, 1C3<br/>
+
-
• Preserve: 1K3, RFP+YFP+1C3.
+
-
 
+
-
</td>
+
-
  </tr>
+
-
</table>
+
-
</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td id="sheetleft">Sept.25th
+
-
Sunday
+
-
 
+
-
</td>
+
-
    <td><table width="620" border="0" cellspacing="0" cellpadding="1">
+
-
      <tr>
+
-
        <td width="244">• Miniprep: 20J, 20H, RFP+YFP+CFP+1K3
+
-
1A3
+
-
</td>
+
-
<td width="164">• Run the gel of PCR results
+
-
 
+
-
</td>
+
-
<td>• Cut RFP+YFP+CFP+1K3
+
-
with E+P<br/>
+
-
• Colony PCR: 13K+10I+1C3
+
-
</td>
+
-
<td>• PCR: fdhF+rush+Vgb+tetR<br/>
+
-
• Preserve: 20H, 20J, 22B
+
-
</td>
+
-
<td>• Run the gel: 20H, 20J, 22B<br/>
+
-
• Cut the bands and purification.
+
-
</td>
+
-
<td>• Run the gel: 13K+10I+1C3, RFP+YFP+CFP+1K3<br/>
+
-
• Culture: 13K+10I+1C3<br/>
+
-
• Grad PCR: fdhF.
+
-
</td>
+
-
      </tr>
+
-
    </table></td>
+
-
  </tr>
+
-
 
+
-
</table>
+
-
 
+
-
</div>
+
-
<div class="block" id="summ">
+
-
<h3 style="color:white;">Data&amp;protocol</h3>
+
-
<hr/>
+
-
<div class="block" id="monday">
+
-
<h3>Monday</h3>
+
-
<hr/>
+
-
<p>WEEK12</p>
+
-
</div>
+
-
<div class="block" id="tuesday">
+
-
<h3>Tuesday</h3>
+
-
<hr/>  
+
-
<p>bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb</p></div>
+
-
<p>&nbsp;</p>
+
-
</div>
+
-
</div>
+
-
<!-- 12 12 12 12 12 12 12 12 12 12 12  -->
+
-
<div class="inner" id="afweek12">
+
-
<div class="block" id="summ">
+
-
<h3 style="color:white;">Data&amp;protocol</h3>
+
-
<hr/>
+
-
<div class="block" id="monday">
+
-
<h3>Monday</h3>
+
-
<hr/>
+
-
<p>WEEK13</p>
+
-
</div>
+
-
<div class="block" id="tuesday">
+
-
<h3>Tuesday</h3>
+
-
<hr/>
+
-
<p>bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb</p></div>
+
-
<p>&nbsp;</p>
+
-
</div>
+
-
</div>
+
-
<!-- 13 13 13 13 13 13 13 13 13 13 13 13 -->
+
-
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+
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Revision as of 16:53, 2 October 2011

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Modeling|biobrick

 Introduction | Model | Simulation 

This model is used for simulating the behavior of three genetic circuits we designed

Introduction

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Retrieved from "http://2011.igem.org/Team:ZJU-China/Notebook"