Preparing chemically competent cells

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*Remove supernatant and replace with 10% original volume TSS.
*Remove supernatant and replace with 10% original volume TSS.
*Make 100uL aliquots and store at <math>-80^o</math>C.
*Make 100uL aliquots and store at <math>-80^o</math>C.
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TSS (50 mL)
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*5g PEG 8000
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*1.5 mL 1M MgCl2
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*2.5 mL DMSO
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*LB to 50 mL
[[Category:E.Coli Protocol]]
[[Category:E.Coli Protocol]]

Revision as of 08:31, 26 May 2005

  • Grow a 5-25mL culture to an OD600 of 0.2 \rightarrow 0.5
  • Centrifuge for 10 minutes at 3000rpm and 4oC.
  • Remove supernatant and replace with 10% original volume TSS.
  • Make 100uL aliquots and store at − 80oC.


TSS (50 mL)

  • 5g PEG 8000
  • 1.5 mL 1M MgCl2
  • 2.5 mL DMSO
  • LB to 50 mL
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