User:Jorge Arturo Zepeda/Notebook/iGem LCG-UNAM team 2010/2010/06/19
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Saturday 19th June 2010I performed two ligations: cI inverter plasmid to GFP(with RBS) and Plasmid 18 to cI inverter PCR and GFP (with RBS), for this fist I had to check in an agarose gel the quantity of the DNA of each part, there are two approaches to ligate, one is to put equimolar quantities of all the parts, and the other is to put the small and the big parts in a ratio of 3:1. The gel came out as follows: 1) Ladder
3) With more or less equimolar concentrations cI-PCR (E/S).................2ul I left the ligations for 10 minutes at room temperature, then to inactivate the enzyme I gave them a heat shock at 62ºC for another 10 minutes.
Those were left in the incubator at 37ºC overnight.
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