Difference: ProtocolsElectrocompetentCells (15 vs. 16)

Revision 162018-10-26 - KateElston

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META TOPICPARENT name="ProtocolList"

Electrocompetent _E. coli

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  1. Pellet the cells by centrifugation for 5 minutes at 6,000 RPM. Remove promptly and pour off supernatant.
  2. Wash by adding 10 ml of chilled 10% glycerol to each tube, then vortexing vigorously to resuspend the pellet. Centrifuge for 3.5 minutes. Remove promptly and pour off supernatant. Repeat for at least four wash cycles in 10% glycerol.
  3. Resuspend in approximately 100 μl of 10% glycerol to make a 100x concentration of the initial culture.
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  1. Divide into 30-50 μl aliquots in 0.5 or 1.7 ml tubes. Freeze or proceed directly to electroporation. Protocol for Electroporation of your Electrocompetent Cells can be found here.
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  1. Divide into 30-50 μl aliquots in 0.5 or 1.7 ml tubes. Freeze or proceed directly to electroporation.
 

Notes

 
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