User:DavidC/22 September 2009

Ligation
Ligation schemes: plasmid / insert:

BBa_B0014 / BBa_P1003 ; BBa_B0014 / BBa_C0012.

First report: Plasmid = 2µL Insert = 3µL

Second report: Plasmid = 1µL Insert = 2µL

Third report: Plasmide = 1µL Insert = 4µL

TAKARA DNA ligation kit
First and third report: Solution A (Buffer) = 25µL Solution B (Enzyme) = 5µL

Second report: Solution A (Buffer) = 15µL Solution B (Enzyme) = 3µL

Incubation 1h at 16°C.

Electroporation
Electroporation cuvettes = 2mm ; inoculums of electrocompetent E.coli DH5alpha= 40µL; pulse = 2,5KVolt ; 1h of incubation.

Spread 1mL of inoculums into a petri dish with LB + ampicillin (50mg/mL) (20/0,02mL).