Erickson: Electroporation

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(New page: Transformation by Electroporation Competent cells Prepare 5 mL of bacterial strain that you wish to transform in BHI. Grow overnight . Chill sterile microfuge tubes on ice and turn on the ...)
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Revision as of 11:57, 2 June 2008

Transformation by Electroporation Competent cells Prepare 5 mL of bacterial strain that you wish to transform in BHI. Grow overnight . Chill sterile microfuge tubes on ice and turn on the refridgerated centrifuge. Add 1 ½ mL of the liquid culture to each microfuge tube until the culture is gone. Keep on ice. Centrifuge for 2-3 minutes at 13,200 rpm. Remove supernatant completely. Add 1 mL COLD glycerol to each tube. Resuspend pellet entirely. Centrifuge again. Remove supernatant and resuspend in 0.5 mL glycerol Remove supernatant and resuspend in 0.25 mL glycerol. Remove supernatant and resuspend in 0.25 mL glycerol. Remove supernatant and resuspend in 100 µL glycerol, combining all tubes into one (resuspend one pellet and use that solution to resuspend the next). Centrifuge. Resuspend pellet in 25µL glycerol. Keep on ice until you add DNA for the transformation.

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