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Electrocompetent cells: Difference between revisions
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*Centrifuge 3 (water): Harvest the cells by centrifugation at 1000g for 20 minutes at 4°C. Decant the supernatant and resuspend the cell pellet in 10 ml ice-cold 10% glycerol. | *Centrifuge 3 (water): Harvest the cells by centrifugation at 1000g for 20 minutes at 4°C. Decant the supernatant and resuspend the cell pellet in 10 ml ice-cold 10% glycerol. | ||
**To spin down your pellet in 10 ml, it might be helpful to use 15-ml Falcon tubes instead of the round-bottom tubes. | |||
*Centrifuge 4 (glycerol): Harvest the cells by centrifugation at 1000g for 20 minutes at 4°C. Carefully decant the supernatant and use a Pastteur pipette attached to a vacuum line to remove any remaining drops of buffer. | *Centrifuge 4 (glycerol): Harvest the cells by centrifugation at 1000g for 20 minutes at 4°C. Carefully decant the supernatant and use a Pastteur pipette attached to a vacuum line to remove any remaining drops of buffer. | ||
