Difference between revisions of "Preparing chemically competent cells"

From OpenWetWare
Jump to: navigation, search
Line 1: Line 1:
 
*Grow a 5-25mL culture to an OD<sub>600</sub> of 0.2 <math>\rightarrow</math> 0.5
 
*Grow a 5-25mL culture to an OD<sub>600</sub> of 0.2 <math>\rightarrow</math> 0.5
 
*Centrifuge for 10 minutes at 3000rpm and <math>4^o</math>C.
 
*Centrifuge for 10 minutes at 3000rpm and <math>4^o</math>C.
*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.
  

Revision as of 06:37, 26 May 2005

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


TSS (50 mL)

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