Electroporation

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Electroporation

In order to transform electro competent cells, a specific protocol must be followed:

1. Chill cuvettes for 1 hour at 4 degrees Celsius prior to performing this transformation.
2. Mix 1uL of DNA into the electrocompetent cells aliquot after the aliquot has thawed in ice.
3. Pipet 55uL of the solution and put it in between metal walls in the cuvette.
4. Turn electrocution rig on. Settings should be Bacteria and MS.
5. Put cuvette in the electrocution rig, and hit pulse. Hold pulse until you hear a beep, at which point, you should immediately let go.
6. The reading should be anywhere from 3-8
7. Take out the cuvette and mix 450uL of SOC media directly in the cuvette. Carefully mix the cells with the SOC media by pipetting up and down.
8. Once the solution is mixed, pipet all the solution and place it in a 1.5mL tube.
9. Incubate for an hour at 37 degrees Celsius
10. Plate 200uL of this mixture