Team:Evry/Notebook/Transposons/13.10.14

BBa_1413044 retrieval of the transposon plasmid

strategy:

Digestion of BBa_1413044 by BglII :
50 ul final volume, 5 ul Buffer NeB 3.1, 5 ul (20 ng/ul) BBa_1413044 in pSB1C3, 1 ul BglII, 39 ul H2O.
Gel electrophoresis:

Purification sur gel:
Kit thermoscientific "Gel extraction". Elution final in 30 ul.

Obtaining the Kan gene on pNK2

PCR: enzyme: Q5; template: 150 ng pNK2 first three well from new stock, next ones the old stock; oligo: F 82/ R 83; Tm tested: 55/60/65; elongation time: 1m30s

Purification sur gel:
Kit thermoscientific "Gel extraction". Elution final in 30 ul.

Oct 13