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From 2010.igem.org

Electroporation

Pipette the DNA samples (5 ng - 2 ~tg in a volume - 3 p.l) to be electroporated into sterile 1.5 ml microfuge tubes. Thaw the competent cells at room temperature for several minutes. Add 50/al of cells to each DNA sample; gently pipette up and down to mix.

Incubate the samples at room temperature for 30 min. ~=.

Set the MicroPulser to "StA". See Section 4 for operating instructions.

Transfer the mixture of cells and D/~IA to a 0.2 cm electroporation cuvette and tap the suspension to the bottom of the tube. Place the cuvette in the chamber slide. Push the slide into the chamber until the cuvette is seated between the contacts in the base of the chamber.

8o Pulse once.

Remove the cuvette from the chamber and immediately add t ml of SMMP medium containing a subinhibitory concentration of antibiotic; gently transfer the cells to a sterile ! 7 x 100 mm tube using a Pasteur pipette. Incubate 1 hr at 37 °C, shaking at 250 rpm.

Check and record the pulse parameters. The time constant should be close to 2.5 milliseconds. The field stren~-rda can be calculated as actual volts (kV) / cuvette gap (cm).

Plate aliquots of the electroporated cells on trypticase soy agar containing selective antibiotic. Incubate plates for 36-48 hrs at 37 °C.