Sauer:Electrocompetent cells

For a nice background on electroporation, visit Wikipedia's page.

Preparing electrocompetent cells:
 * 1) Make a 1/100X dilution of your overnight culture into fresh medium (and add antibiotics if desired).
 * 2) Grow your culture to an OD600 nm ~ 0.5-0.7.
 * 3) Meanwhile, chill sterile water and a sterile 10% glycerol solution at 4&deg;C.
 * 4) For a 1 L culture I'd chill 500 mL of water and 100 mL of 10% glycerol, and this is more than enough.:Marylee 23:04, 27 November 2006 (EST)
 * 5) Place your cultures over ice, ~15'.
 * 6) Spin at 4000 rpm, 15', and dump the supernatant.
 * 7) Gently resuspend the pellet in 1 mL of chilled, sterile water.  Transfer to a 50 mL Falcon tube and fill it with more water.
 * 8) Spin at 4000 rpm, 15', and dump the supernatant.
 * 9) Repeat this resuspension-spin cycle four times, the final time using your 10% glycerol solution.
 * 10) Resuspend your pellet in 500 &mu;L of 10% glycerol.
 * 11) Aliquot 60 &mu;L into eppendorf tubes and freeze at -80&deg;C until use.

Electroporation:
 * 1) Thaw your aliquot of electrocompetent cells on ice and transfer them to an electroporation cuvette.
 * 2) Add 1 &mu;L of DNA, or whatever else your introducing, to your cells.
 * 3) Electroporate according to the specifications of your equipment and sample.
 * 4) Quickly add 1 mL of medium (lacking antibiotics) to the cuvette and then transfer to an eppendorf.
 * 5) Cure cells by shaking at 37&deg;C (or whatever temperature you would use to plate your cells) for ~1 hour.
 * 6) Gently pellet cells by spinning at 4000 rpm, 3'.
 * 7) Resuspend the pellet in roughly 150 uL of supernatant and plate on selective medium.