Team:KIT-Kyoto/Notebook-week2p

From 2012.igem.org

(Difference between revisions)
Line 70: Line 70:
                 <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>
-
                <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 31st


 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  25 cycle 
  68℃  30sec 
  14℃  ∞ 


September 1st




 All samples 
 DNA template  0.3uL 
 10× KOD plus buffer  10uL 
 2mM dNTPs  10uL 
 25mM MgSO4  3.2uL 
 10P 5’ primer  3uL 
 10P 3’ primer  3uL 
 KOD plus  2uL 
 dH2O  68.5uL 
 Total  100uL 


 Temperature  Time  Circle 
  95℃  2min 
  95℃  15sec  25 cycle 
  59℃  30sec  25 cycle 
  68℃  1min30sec(Act5c and LacZ) or 2min(GAL4 and U-T)  25 cycle 
  68℃  1min30sec(Act5c and LacZ) or 2min(GAL4 and U-T) 
  14℃  ∞ 




September 3rd


 All samples 
 DNA template  0.2uL 
 10× KOD plus buffer  5uL 
 2mM dNTPs  5uL 
 25mM MgSO4  1.6uL 
 10P 5’ primer  1.5uL 
 10P 3’ primer  1.5uL 
 KOD plus  1u L
 dH2O  34.2uL 
 Total  50uL 


 Temperature  Time  Circle 
 95℃  2min 
 95℃  15sec  25 cycle 
 55℃(GAL4) and 58℃(Except for GAL4)  30sec  25 cycle 
 68℃  2min30sec  25 cycle 
 68℃  2min30sec 
 14℃  ∞ 




September 4th


 9/3 GAL4  40uL 
 M buffer(TOYOBO)  5uL 
 XbaⅠ(TOYOBO)  0.5uL 
 SpeⅠ(TOYOBO)  0.5uL 
 dH2O  4uL 
 Total  50uL 


 each DNA  40uL 
 3 buffer(NEB)  5uL 
 BglⅡ(NEB)  0.5uL 
 dH2O  4.5uL 
 Total  50uL 


 9/3 GAL4  23uL 
 M buffer(TOYOBO)  10uL 
 XbaⅠ(TOYOBO)  1uL 
 SpeⅠ(TOYOBO)  1uL 
 CIP  0.5uL 
 dH2O  64.5uL 
 Total  100uL 




September 5th


 EGFP  pSB1C3 
 DNA template  40uL  23uL 
 3 buffer  5uL  5uL 
 BglⅡ  0.5uL  0.5uL 
 dH2O  4.5uL  21.5uL 
 Total  50uL  50uL 




 DNA template  40uL 
 4 buffer(NEB)  5uL 
 SpeⅠ  0.5uL 
 100×BSA  0.5uL 
 CIP  0.5uL 
 dH2O  4uL 
 Total  50uL 


 1uL 
 1uL 
 dH2O  3uL 
 Ligation high  2.5uL 
 Total  7.5uL