Electroporation Protocol
Turn on electroporator and set to 1.7 - 2.5 kv ( optimize for strain ) , 200 ohms and 25 µF .
Place recovery SOC in 37°C water bath .
Pre - warm LB - antibiotic plates at 37°C .
Thaw cells on ice for 10 min or use freshly made cells .
Place appropriate number of microcentrifuge tubes and 1 mm - electroporation cuvettes on ice .
Flick the tube containing cells a few times to mix and add 25 µl to the microcentrifuge tubes .
Add 1 µl of a 10 pg / µl DNA solution ( in DI water ) to the cells in the microcentrifuge tube .
Transfer the DNA - cell mixture to the cold cuvette , tap on countertop 2X , wipe water from exterior of cuvette and place in the electroporation module and press pulse ( don’t hold the button down ) .
Immediately add 975 µl of 37°C SOC , mix by pipetting up and down once and transfer to a 15 ml - falcon tube .
Rotate in the 37°C incubator for 1 h .
Make appropriate dilutions .
Incubate overnight at 37°C .
