Electrotransformation of plasmids in E. coli
- Grow 2 ml of the bacteria host overnight in LB medium
- OPTIONAL: On the next morning, transfer the culture to a microtube and heat shock the culture for exactly 15 min at 42ºC
- Chill the culture in ice slurry for 10 min. For now on, all procedures are made at 4ºC
- Harvest cells at 10,000 rpm for 3 min. Resuspend in 1 ml of ice-cold and centrifuge again. Repeat this step twice more.
- Resuspend bacteria in a final volume of 50 μl in ice cold
- Electroporation is made in a prechilled 0.2 ml cuvette in the Bio-Rad electroporator according to the manufacturer instructions.
- Following electroporation, immediately add 2 ml of SOC (2% tryptone, 0.5% yeast extract, 10 mM NaCl, 10 mM , 10 mM ) or LB.
- Cells are transferred to a tube and incubated at 37ºC for at least 1h to allow expression of the AbR phenotype.
- Plate 0.1-2 ml bacteria on the appropriate antibiotic plates and incubate O.N. at 37ºC.