Adapted from: http://openwetware.org/wiki/Sauer:Electrocompetent_cells
This is for transforming E. coli
- Remove cells from -80 and chill on ice for 10 minutes or until the culture is thawed
- Meanwhile: label 1 1.5 microcentrifuge tube per transformation and place aside
- Add 1 ul of plasmid DNA or __ ul of ligation to the competent cells
- Flick to mix
- Transfer to a fresh electroporation cuvette using a pipette
- Tap the tube on the bench to make sure all the culture reaches the bottom of the cuvette
- Electroporate based on the machine's specs
- For E. coli in 1 mm cuvettes- use the EC1 program
- IMMEDIATELY ADD 1 ML OF SOC TO THE CUVETTE AND TRANSFER TO A 1.5 ml MICROCENTRIFUGE TUBE USING THE INCLUDED PASTEUR PIPETTE
- THIS IS ESSENTIAL. 3X DROP IN EFFICIENCY PER MINUTE WAITED TO ADD SOC
- Shake the 1.5 ml centrifuge tube at the appropriate temperature for 1 hr.
- Pellet the cells by centrifuging at 4000 rpm for 3 min
- Gently dump the supernatant except for 150 ul
- Resuspend the pellet in the remaining 150 ul
- Plate on the appropriate plates