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Team:Evry/Notebook/Transposons/16.10.14
From 2014.igem.org
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| - | + | <h4> defining where the transposase integrate </h4> | |
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find enzyme with a good cut numbers in the genome and not in our insert: HindIII | find enzyme with a good cut numbers in the genome and not in our insert: HindIII | ||
Revision as of 22:13, 17 October 2014
Show BBa_1413044 work even empty in DH5alpha
8 colonies of DH5alpha were transformed with BBa_1413044 and grown in 3 ml LB + 2 negative control
Preparation for PCR colony:
Spin down 500 ul culture at 14000 g - 2 min, resuspend in 100 ul H20 MilliQ, put at 95 C° for 10 min then spin down at 14000 g - 2 min.
PCR: enzyme: Q5; template: 3 ul of supernatent for the culture; oligo: F 100/ R 101; Tm tested: 55; elongation time: 1m00s
defining where the transposase integrate
find enzyme with a good cut numbers in the genome and not in our insert: HindIII
Then religate with T4
then light digestion with XbaI
then PCR
Aug 08
