Team:Hong Kong HKUST/notebook/mod3

From 2013.igem.org

(Difference between revisions)
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       <div>
       <div>
       <a href='#' class='head'>Week 4</a>
       <a href='#' class='head'>Week 4</a>
-
       <div class='content' align="left">Wet lab<br>
+
       <div class='content' align="left">
-
·      Inoculation of pBlueScript KS(+) for training<br>
+
<p>&nbsp;</p>
-
·      Autoclave basic materials<Br>
+
<p><strong>24-6-2013</strong> <br />
-
·      Preparing LB<br>
+
  Inoculation of pcmv/myc/mito </p>
-
·      LB-Ampicillin plates poured<br>
+
<p>&nbsp;</p>
-
Dry lab<br>
+
<p><strong>25-6-2013</strong> <br />
-
Protocols review
+
  Plasmid extraction for pcmv/myc/mito <br />
 +
  Digestion of pcmv plasmid extraction product with PstI-HF and NotI-HF <br />
 +
  Gel electrophoresis of pcmv digested product <br />
 +
Gel check for pcmv digestion product </p>
 +
<p>&nbsp;</p>
 +
<p><strong>26-6-2013</strong> <br />
 +
  Gel purification of pcmv gel product <br />
 +
  Gel check of pcmv gel purified product <br />
 +
  PCR clone MLS from pcmv/myc/mito <br />
 +
Transformation of pEGFP-N1 with SURE </p>
 +
<p>&nbsp;</p>
 +
<p><strong>27-6-2013</strong> <br />
 +
  Gel check for the MLS PCR product and PCR Clean Up <br />
 +
  Digestion of MLS clean up product with EcoRI &amp; PstI-HF <br />
 +
Inoculation of pEGFP-N1 </p>
 +
<p>&nbsp;</p>
 +
<p><strong>28-6-2013</strong> <br />
 +
  PCR Clean up of digested MLS <br />
 +
  Plasmid Extraction of pEGFP-N1 <br />
 +
Digestion of pEGFP-N1 with PstI-HF  &amp; NotI-HF </p>
 +
<p>&nbsp;</p>
       </div>
       </div>
       </div>
       </div>
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   <div>
   <div>
       <a href='#' class='head'>Week 1</a>
       <a href='#' class='head'>Week 1</a>
-
       <div class='content' align="left">Content 1
+
       <div class='content' align="left">
 +
<p>&nbsp;</p>
 +
<p><strong>02-07-2013</strong> <br />
 +
  Gel Check of pEGFP-N1 plasmid extraction product to confirm presence of DNA <br />
 +
  Digestion of pEGFP-N1 (with NotI-HF and PstI-HF) 2nd EGFP product:repeat digestion <br />
 +
  Inoculation of pEGFP-N1 3rd EGFP: repeat Inoculation <br />
 +
  Get pSB1C3.J04450 plasmid extraction product from Bida <br />
 +
  Digestion of pSB1C3 with EcoRI and PstI-HF </p>
 +
<p>&nbsp;</p>
 +
<p><strong>03-07-2013</strong> <br />
 +
  Gel electrophoresis of digested EGFP (2nd) <br />
 +
  Plasmid extraction of pEGFP-N1 (3rd) <br />
 +
  Digetion of pEGFP-N1 (3rd) using NotI-HF and PstI-HF <br />
 +
  Ligation of MLS with pSB1C3 <br />
 +
Transformation of ligated pSB1C3-MLS </p>
 +
<p>&nbsp;</p>
 +
<p><strong>04-07-2013</strong> <br />
 +
  Gel purification of pEGFP-N1 (2nd) (cont.) <br />
 +
  Gel analysis and gel electrophoresis of pEGFP-N1 (3rd) <br />
 +
  Run gel for remaining 3rd EGFP (digested) <br />
 +
  Ethanol precipitation of pEGFP-N1 (4th) (using remaining 3rd EGFP plasmid) <br />
 +
  Digestion of pEGFP-N1 (4th) and gel electrophoresis <br />
 +
  Transformation of pEGFP-N1 into DH10B <br />
 +
  Colony PCR of MLS biobrick and gel electrophoresis <br />
 +
Inoculation of MLS colony no.2 &amp; spread the remaining re-suspend colony 2 on CAM plate </p>
 +
<p>&nbsp;</p>
 +
<p><strong>05-07-2013</strong> <br />
 +
  pEGFP-N1 troubleshoot <br />
 +
  Digestion by PstI-HF, NotI-HF, PstI-HF +NotI-HF and gel check <br />
 +
  Inoculate EGFP from transformed new competent cell <br />
 +
  Plasmid extraction of MLS Biobrick <br />
 +
Sanger sequencing of MLS </p>
 +
<p>&nbsp;</p>
       </div>
       </div>
   </div>
   </div>
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   <div>
   <div>
       <a href='#' class='head'>Week 2</a>
       <a href='#' class='head'>Week 2</a>
-
       <div class='content' align="left">Content 1
+
       <div class='content' align="left">
       </div>
       </div>
   </div>
   </div>
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       <a href='#' class='head'>Week 1</a>
       <a href='#' class='head'>Week 1</a>
       <div class='content' align="left">
       <div class='content' align="left">
-
<p>&nbsp;</p>
 
-
<p><strong>August 5</strong></p>
 
-
<p>GFP (25)PCR Check with VR VF2</p>
 
-
<ul>
 
-
  <blockquote>
 
-
    <p>- Gel check(overan)</p>
 
-
  </blockquote>
 
-
</ul>
 
-
<p>BBa_J176171</p>
 
-
<ul>
 
-
  <blockquote>
 
-
    <p>- Digestion using SpeI and PstI-HF to extract the backbone</p>
 
-
  </blockquote>
 
-
</ul>
 
-
<p>PolyA / BBa_K404108</p>
 
-
<ul>
 
-
  <blockquote>
 
-
    <p>- Digestion using XbaI and PstI-HF</p>
 
-
    <p>- Gel electrophoresis</p>
 
-
    <p>- Run Gel </p>
 
-
  </blockquote>
 
-
</ul>
 
-
<p>Inoculation of pSB1C3</p>
 
-
<p>&nbsp;</p>
 
-
<p><strong>August 6</strong></p>
 
-
<p>GFP (25)</p>
 
-
<ul>
 
-
  <blockquote>
 
-
    <p>- Repeat PCR check</p>
 
-
    <p>- Run Gel</p>
 
-
  </blockquote>
 
-
</ul>
 
-
<p>pSB1C3</p>
 
-
<ul>
 
-
  <blockquote>
 
-
    <p>- Plasmid extraction</p>
 
-
  </blockquote>
 
-
</ul>
 
-
<p>PolyA / BBa_K404108</p>
 
-
<ul>
 
-
  <blockquote>
 
-
    <p>- Gel purification</p>
 
-
    <p>- Gel check</p>
 
-
  </blockquote>
 
-
</ul>
 
-
<p>BBa_J176171</p>
 
-
<ul>
 
-
  <blockquote>
 
-
    <p>- PCR clean up</p>
 
-
  </blockquote>
 
-
</ul>
 
-
<p>BBa_K929000 &amp; BBa_J52034 </p>
 
-
<ul>
 
-
  <blockquote>
 
-
    <p>- Transformation using CAM plate</p>
 
-
  </blockquote>
 
-
</ul>
 
-
<p>MLS</p>
 
-
<ul>
 
-
  <blockquote>
 
-
    <p>- PCR amplify MLS in RFC 10 and 25 format</p>
 
-
  </blockquote>
 
-
</ul>
 
-
<p>&nbsp;</p>
 
-
<p><strong>August 7</strong></p>
 
-
<p>MLS (RFC 10 &amp; 25)</p>
 
-
<ul>
 
-
  <blockquote>
 
-
    <p>- PCR clean up</p>
 
-
    <p>- Nanodrop for the PCR clean up product</p>
 
-
    <p>- Gel check for the PCR clean up product</p>
 
-
    <p>- Digestion using EcoRI and PstI</p>
 
-
  </blockquote>
 
-
</ul>
 
-
<p>PolyA / BBa_K404108</p>
 
-
<ul>
 
-
  <blockquote>
 
-
    <p>- Gel check</p>
 
-
  </blockquote>
 
-
</ul>
 
-
<p>pSB1C3 </p>
 
-
<ul>
 
-
  <blockquote>
 
-
    <p>- Digest with EcoRI &amp; PstI </p>
 
-
    <p>- Run gel and cut gel </p>
 
-
  </blockquote>
 
-
</ul>
 
-
<p>GFP(25) </p>
 
-
<ul>
 
-
  <blockquote>
 
-
    <p>- Digestion with EcoRI &amp; SpeI</p>
 
-
    <p>- Run gel and cut gel </p>
 
-
  </blockquote>
 
-
</ul>
 
-
<p>&nbsp;</p>
 
-
<p><strong>August 8</strong></p>
 
-
<p>GFP25 &amp; pSB1C3</p>
 
-
<ul>
 
-
<blockquote>
 
-
  <p>- Gel purification</p>
 
-
    <p>- Nanodrop</p>
 
-
    <p>- Gel check </p>
 
-
  </blockquote>
 
-
</ul>
 
-
<p>polyA</p>
 
-
<ul>
 
-
  <blockquote>
 
-
    <p>- Digestion with EcoRI and XbaI</p>
 
-
    <p>- PCR clean up</p>
 
-
  </blockquote>
 
-
</ul>
 
-
<p>BBa_K929000 &amp; BBa_J52034 </p>
 
-
<ul>
 
-
  <blockquote>
 
-
    <p>- Inoculation </p>
 
-
  </blockquote>
 
-
</ul>
 
-
<p>&nbsp;</p>
 
-
<p><strong>August 9</strong></p>
 
-
<p>BBa_K929000 &amp; BBa_J52034 </p>
 
-
<p>- Plasmid extraction: J52034 is not transformed</p>
 
-
<p>MLS.pSB1C3 &amp; GFP.PolyA</p>
 
-
<p>- Ligation</p>
 
-
<p>- Transformation to SURE on CAM plate</p>
 
-
<p>&nbsp;</p>
 
-
 
       </div>
       </div>
     </div>
     </div>

Revision as of 18:09, 19 September 2013








June 2013

Week 4

 

24-6-2013
Inoculation of pcmv/myc/mito

 

25-6-2013
Plasmid extraction for pcmv/myc/mito
Digestion of pcmv plasmid extraction product with PstI-HF and NotI-HF
Gel electrophoresis of pcmv digested product
Gel check for pcmv digestion product

 

26-6-2013
Gel purification of pcmv gel product
Gel check of pcmv gel purified product
PCR clone MLS from pcmv/myc/mito
Transformation of pEGFP-N1 with SURE

 

27-6-2013
Gel check for the MLS PCR product and PCR Clean Up
Digestion of MLS clean up product with EcoRI & PstI-HF
Inoculation of pEGFP-N1

 

28-6-2013
PCR Clean up of digested MLS
Plasmid Extraction of pEGFP-N1
Digestion of pEGFP-N1 with PstI-HF  & NotI-HF

 

July 2013

Week 1

 

02-07-2013
Gel Check of pEGFP-N1 plasmid extraction product to confirm presence of DNA
Digestion of pEGFP-N1 (with NotI-HF and PstI-HF) 2nd EGFP product:repeat digestion
Inoculation of pEGFP-N1 3rd EGFP: repeat Inoculation
Get pSB1C3.J04450 plasmid extraction product from Bida
Digestion of pSB1C3 with EcoRI and PstI-HF

 

03-07-2013
Gel electrophoresis of digested EGFP (2nd)
Plasmid extraction of pEGFP-N1 (3rd)
Digetion of pEGFP-N1 (3rd) using NotI-HF and PstI-HF
Ligation of MLS with pSB1C3
Transformation of ligated pSB1C3-MLS

 

04-07-2013
Gel purification of pEGFP-N1 (2nd) (cont.)
Gel analysis and gel electrophoresis of pEGFP-N1 (3rd)
Run gel for remaining 3rd EGFP (digested)
Ethanol precipitation of pEGFP-N1 (4th) (using remaining 3rd EGFP plasmid)
Digestion of pEGFP-N1 (4th) and gel electrophoresis
Transformation of pEGFP-N1 into DH10B
Colony PCR of MLS biobrick and gel electrophoresis
Inoculation of MLS colony no.2 & spread the remaining re-suspend colony 2 on CAM plate

 

05-07-2013
pEGFP-N1 troubleshoot
Digestion by PstI-HF, NotI-HF, PstI-HF +NotI-HF and gel check
Inoculate EGFP from transformed new competent cell
Plasmid extraction of MLS Biobrick
Sanger sequencing of MLS

 

Week 2
Week 3
Content 1
Week 4
Content 1
Week 5
Content 1

August 2013

Week 1
Week 2
Content 1
Week 3
Content 1
Week 4
Content 1

September 2013

Week 1
Content 1
Week 2
Content 1
Week 3
Content 1
Week 4
Content 1
Week 5
Content 1

Retrieved from "http://2013.igem.org/Team:Hong_Kong_HKUST/notebook/mod3"