User:DavidC/23 September 2009

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Contents

Wednesday the 23th

Ligation

Ligation scheme: plasmid / insert:

BBa_C0012/BBa_B0014

First report:
Plasmid = 0,5µL
Insert = 3µL

Second report:
Plasmid = 0,5µL
Insert = 7µL

Third report:
Plasmid = 0,5µL
Insert = 1µL

Fourth report:
Plasmid = 0,5µL
Insert = 5µL

NEB Enzymes

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

Ligation mix (NEB):
2µL of T4 ligase + 2µL of 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).

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