User:DavidC/11 September 2009

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framless



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Friday the 11th

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).