User:DavidC/22 September 2009

From 2009.igem.org

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Contents

Tuesday the 22th

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).