Team:Hong Kong HKUST/notebook/mod3
From 2013.igem.org
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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"> | + | <div class='content' align="left"> |
- | + | <p> </p> | |
- | + | <p><strong>24-6-2013</strong> <br /> | |
- | + | Inoculation of pcmv/myc/mito </p> | |
- | + | <p> </p> | |
- | + | <p><strong>25-6-2013</strong> <br /> | |
- | + | 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> </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> </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 & PstI-HF <br /> | ||
+ | Inoculation of pEGFP-N1 </p> | ||
+ | <p> </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 & NotI-HF </p> | ||
+ | <p> </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"> | + | <div class='content' align="left"> |
+ | <p> </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> </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> </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 & spread the remaining re-suspend colony 2 on CAM plate </p> | ||
+ | <p> </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> </p> | ||
</div> | </div> | ||
</div> | </div> | ||
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<a href='#' class='head'>Week 2</a> | <a href='#' class='head'>Week 2</a> | ||
- | <div class='content' align="left"> | + | <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> | ||
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Revision as of 18:09, 19 September 2013
June 2013
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
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