Team:Evry/Notebook/Transposons/16.10.14

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Picture


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