User:DavidC/11 September 2009

Ligation
Ligation pairs plasmid / insert : BBa_B0014/ BBa_P1002 BBa_B0030 / BBa_R0040 BBa_B0014 / BBa_C0012

First report: Plasmid = 1µL Insert = 4µL

Second report: Plasmid = 1µL Insert = 2µL

For each samples add sterilized water to obtain a maximum volume equals to 8µL.

Ligation mix (NEB): 6µL T4 ligase + 6µL T4 ligase buffer. Add 2µL / tube. Incubation 1h at RT.

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).