User:Jorge E. Buendia Buendia/Notebook/iGEM UNAM-Genomics-Mexico/2010/10/11: Difference between revisions
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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. | |||
Revision as of 08:59, 11 October 2010
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October 11th, 20101. Inactivate restriction enzyme for 20 min at 80º C.
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