Difference between revisions of "Preparing chemically competent cells"

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TSS (50 mL)
+
[[TSS]] (50 mL)
 
*5g PEG 8000
 
*5g PEG 8000
 
*1.5 mL 1M MgCl2
 
*1.5 mL 1M MgCl2

Revision as of 07:37, 26 May 2005

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


TSS (50 mL)

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