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.


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