User:Jorge E. Buendia Buendia/Notebook/iGEM UNAM-Genomics-Mexico/2010/10/11

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October 11th, 2010
1. Inactivate restriction enzyme for 20 min at 80º C.

2. Make the following ligations:
 * pSB3K3 + J23101 SpeI-PstI (3ul) with ΔRBS + GFP E004 XbaI-PstI (5ul)
 * pSB4A5 + MinBP SpeI-PstI (3ul) with ΔRBS + GFP E004 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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