Electroporation Procedure

 

Materials:

Methods:

1. Set electroporator to charging voltage ( . Pulse length is fixed at 5-6 msec.
2. Pipet of cell culture into cuvette
3. Pipet of plasmid into cuvette. Mix by pipetting up and down. Can also ``flick'' cuvette. (Note: suspension must touch both side walls of cuvette).
4. Chill cuvette on ice for 1 min
5. Switch Knob A from CHARGE to PULSE
6. Immediately add SOC
7. Briefly and gently pipet up and down to mix thoroughly
8. Transfer to polypropylene tube, and hold for 1 hour (shaking at 225 rpm may improve recovery)
9. Transfer to appropriate agar plate
10. Result: transformants/ DNA