NanoBio: Bacterial Transformation using Electroporation
Simultaneous Transformation using Electroporation
If there are electrocompetent cells available to you in the -80°C, thaw the cells on ice and begin. If you need to make a fresh batch of electrocompetent cells, use either the small-scale or large-scale * Prep for Electrocompetent cells.
- Chill cuvettes for at least 5 minutes on ice.
- Set electroporator to 2.5kV. (Our electroporator is set to 600Ω)
- Add 5pg – 0.5ug DNA to cells. Mix by pipetting. Pippet into a sterile e.p. cuvette.
- Note: This DNA should be salt-free, so when purifying with kit, use water.
- Note: Include enough samples for +/- PCR.
- Note: The DNA should not sit in the cells for more than 1 minute.
- Make sure to dry the cuvette to prevent arcing!
- Place the DRY cuvette into the sample chamber. Apply the pulse by pushing the pulse button twice. Remove the cuvette.
- Note: If you hear a popping sound, this means that the sample has arced. This could either be (1) too much salt in your sample or (2) a wet cuvette.
- Immediately add 1mL of room-temp SOC or LB and transfer to a culture tube
- Incubate 1 hr with shaking at 37°C.
- Plate out cells on LB+antibiotic. Grow o/n at 37°C.
- Heather M. Jensen 19:52, 11 June 2008 (CST):
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