Matt Gethers/CRI, Thailand/Labwork/Electroporation

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Electroporation Protocol

  1. Grow up a 6 ml overnight culture of Pseudomonas in LB.
  2. Spin down the culture at 5,000 RPM for 5 minutes at room temperature.
  3. Wash twice in 1 ml 300 mM sucrose.
  4. Resuspend pellet in 100 μl 300 mM sucrose.
  5. Add the 100 μl to a cuvette (also at room temperature) and pulse at 2.5 kV, 200 Ω, and 20 μF.
  6. Add 1 ml SOC to the cuvette immediately following the pulse.
  7. Spread 100 μl on a plate containing the appropriate antibiotic and incubate at 37 degrees over night.