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Making electro-competent E. coli
LAB PROTOCOL
Day 1: Inoculate LB-medium with E. coli strain to make an overnight culture (5ml)
Day 2: Inoculate fresh LB-medium (30ml) from overnight culture at 1:100 dilution (300µl)
Grow at 37°C to an OD600 0.5-0.7 (for 3-4hrs)
Pellet by centrifuging (4000rpm for 10 minutes or 6500rpm for 5 minutes) and remove supernatant
Wash cells with ice-cold distilled water (30ml) and pellet again. Repeat wash step 3x.
Re-suspend cells in 200µl in 40% glycerol
Aliquot 50µl of cells in PCR tubes and store at -80°C.
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