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Electroporation
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Latest revision as of 02:04, 28 September 2013
Main Page
Electroporation
In order to transform electro competent cells, a specific protocol must be followed:
- Chill cuvettes for 1 hour at 4 degrees Celsius prior to performing this transformation.
- Mix 1uL of DNA into the electrocompetent cells aliquot after the aliquot has thawed in ice.
- Pipet 55uL of the solution and put it in between metal walls in the cuvette.
- Turn electrocution rig on. Settings should be Bacteria and MS.
- Put cuvette in the electrocution rig, and hit pulse. Hold pulse until you hear a beep, at which point, you should immediately let go.
- The reading should be anywhere from 3-8
- 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.
- Once the solution is mixed, pipet all the solution and place it in a 1.5mL tube.
- Incubate for an hour at 37 degrees Celsius
- Plate 200uL of this mixture
