User:DavidC/16 September 2009

DNA extraction
DNA extraction from bio-brick R0040+B0030 and P1002+C0040 from 500mL of E.coli DH5alpha growth in LB medium.

Ligation: samples digested on the 15th of September
Ligation pairs: plasmid / insert

BBa_B0014 / BBa_I712669 ; BBa_P1003/BBa_B0014 ; BBa_J23100/BBa_B0014

First report: Plasmid = 1µL Insert = 6µL

Second report: Plasmid = 3µL Insert = 5µL

Third report: Plasmid = 1µL Insert = 5µL

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

Ligation mix (NEB): 9µL of T4 ligase + 9µL 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).