Matt Gethers/CRI, Thailand/Labwork/Electroporation

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.