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.


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