Team:UNIPV-Pavia/Methods Materials/Ligation

From 2009.igem.org

(Difference between revisions)
(→Ligation)
(→Ligation)
Line 57: Line 57:
*'''10X Roche T4 Ligase Buffer'''
*'''10X Roche T4 Ligase Buffer'''
*'''ddH2O'''
*'''ddH2O'''
-
<br>
 
</td></tr></table></div><br><br><br><br>
</td></tr></table></div><br><br><br><br>
</td></tr></table>
</td></tr></table>

Revision as of 11:57, 30 June 2009

EthanolPVanimation.gif



Protocols

Ligation

(estimated time: 20 min + 12-16 hours overnight incubation)

  • (For every ligation)
  • Add 50 ng of vector
  • Add      
  • Heat DNA mix at 65°C for 5 min for DNA denaturation
  • Add 1 µl of T4 Ligase buffer
  • Add 1 µl of T4 Ligase
  • 10 µl final volume
  • Incubate at 16°C overnight


  • Then, ligation can be conserved at 4°C or can be transformed
  • Before transformation you have to inactivate T4 Ligase:
    • Heat ligation at 65°C for 10 min.






  • Roche T4 Ligase
  • 10X Roche T4 Ligase Buffer
  • ddH2O




Retrieved from "http://2009.igem.org/Team:UNIPV-Pavia/Methods_Materials/Ligation"