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INSA-Lyon/27 July 2010
From 2010.igem.org
July 27
Extraction of L1 in 40µL
Digestion of 5 colonies of L2 by X+P
Gel electrophoresis -->1 colony of L2 has the good insert (L2G)
Digestion of L1 by S+P and E+P and digestion of L2G by X+P
Agarose gel electrophoresis --> good insert for L1 et L2
Ligation L3: 10 µL of L2 , 5µL of L1 in 20µL
Transformation of the ligation 1F/24A
Extraction of DNA: 1F, 24A, 14K
Agarose gel electrophoresis: ok except 14K whose the band do not have the size expected !?
Digestion: 1F by S+P, 24A by X+P
Agarose gel electrophoresis: 1F-->oK,24A: not digested (we forgot to centrifugate after adding the enzymes)
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