User:Jorge E. Buendia Buendia/Notebook/iGEM UNAM-Genomics-Mexico/2010/09/10

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September 10th, 2010

1. Replate transformation of BBa_I20260 Δ RBS into LB solid medium with Kan 30 because cells plated on September 9th, didn’t grow, 36 colonies were plated.

2. Make ligation of BBa_I20260 with the new RBS.

  • Ligation methods (Total volume 20ul):
DNA -> 7ul
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.