User:Jorge E. Buendia Buendia/Notebook/iGEM UNAM-Genomics-Mexico/2010/09/19

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September 19th, 2010

1. Make the following ligations:

  • pSB3K3-J23101 SpeI-PstI (3ul) with Δ RBS-GFP E0040 XbaI-PstI (5ul).
  • pSB3K3-Min. Blue Promoter SpeI-PstI (2ul) with Δ RBS-GFP E0040 XbaI-PstI (5ul).
  • Ligation methods (Total volume 20ul):
DNA -> Volume of each part to be ligated is indicated above.
Buffer for T4 DNA ligase 5X -> 4ul (Final concentration 20%)
T4 DNA ligase -> 1ul
HPLC -> Complete total volume (20ul)
Incubate overnight at 16ºC
Mix well (vortex) buffer and reaction tubes, unfreeze buffer on ice.