Team:KIT-Kyoto/Notebook-week1p

From 2012.igem.org

(Difference between revisions)
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                 <li><a href="http://2012.igem.org/Team:KIT-Kyoto/Notebook-week4p"><img src="http://2012.igem.org/wiki/images/3/31/Side_week4kit.jpg" width="150" height="30"></a></li>
                 <li><a href="http://2012.igem.org/Team:KIT-Kyoto/Notebook-week4p"><img src="http://2012.igem.org/wiki/images/3/31/Side_week4kit.jpg" width="150" height="30"></a></li>
                 <li><a href="http://2012.igem.org/Team:KIT-Kyoto/Notebook-week5p"><img src="http://2012.igem.org/wiki/images/e/e7/Side_week5kit.jpg" width="150" height="30"></a></li>
                 <li><a href="http://2012.igem.org/Team:KIT-Kyoto/Notebook-week5p"><img src="http://2012.igem.org/wiki/images/e/e7/Side_week5kit.jpg" width="150" height="30"></a></li>
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                <li><a href="http://2012.igem.org/Team:KIT-Kyoto/Notebook-week6p"><img src="http://2012.igem.org/wiki/images/e/e8/Side_week6kit.jpg" width="150" height="30"></a></li>
 
             </ul>
             </ul>
         </li>
         </li>

Revision as of 10:22, 25 September 2012






August 25th


 All DNA  500ng  1ug 
 pEGFP-C  3uL  6uL 
 H buffer(TOYOBO)  5uL  5uL 
 BamHⅠ(TOYOBO)  1uL  1uL 
 BglⅡ(TOYOBO)  1uL  1uL 
 100×BSA  0.5uL  0.5uL 
 dH2O  39.5uL  36.5uL 
 Total  50uL  50uL 




 DNA sample  23uL 
 4 buffer(NEB)  5uL 
 EcoRⅠ-HF(NEB)  1uL 
 SpeⅠ(NEB)  1.5uL 
 100×BSA  0.5uL 
 dH2O  19uL 
 Total  50uL 


 All samples 
 DNA template  0.5uL 
 10× KOD plus buffer  10uL 
 2mM dNTPs  10uL 
 25mM MgSO4  3.2uL 
 10P 5’ primer  3uL 
 10P 3’ primer  3uL 
 KOD plus  2uL 
 dH2O  68.3uL 
 Total  100uL 


 Temperature  Time  Circle 
  95℃ 2min
  95℃ 15sec 25 cycle 
  58℃ 30min(UAS) or 2min10sec(pSB1C3) 25 cycle 
  68℃ 2min30sec 25 cycle 
  68℃ 2min30sec
  14℃ ∞


August 26th




August 27th


 pEGFP-C2(cut)  1uL 
 Ligation high  2.5uL 
 dH2O  5uL 
 Total  7.5uL 






August 28th


August 29th


 pEGFP-C2-1, -2, -3, -4  1uL 
 10×H buffer(TOYOBO)  0.5uL 
 XhoⅠ  0.1uL 
 PstⅠ  0.1uL 
 dH2O  3.3uL 
 Total  5uL 




 All samples 
 DNA template  0.5uL 
 10× KOD plus buffer  10uL 
 2mM dNTPs  10uL 
 25mM MgSO4  3.2uL 
 10P 5’ primer  3uL 
 10P 3’ primer  3uL 
 KOD plus  2uL 
 dH2O  68.3uL 
 Total  100uL 


 Temperature  Time  Cycle 
 95℃  2min 
 95℃  15sec  25 cycle 
 58℃  30sec  25 cycle 
 68℃  30sec(UAS) or 1min(EGFP)  25 cycle 
 68℃  30sec(UAS) or 1min(EGFP) 
 14℃  ∞