Preparing chemically competent cells: Difference between revisions
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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. | ||
TSS (50 mL) | |||
*5g PEG 8000 | |||
*1.5 mL 1M MgCl2 | |||
*2.5 mL DMSO | |||
*LB to 50 mL | |||
[[Category:E.Coli Protocol]] | [[Category:E.Coli Protocol]] | ||
Revision as of 06:31, 26 May 2005
- Grow a 5-25mL culture to an OD600 of 0.2 [math]\displaystyle{ \rightarrow }[/math] 0.5
- Centrifuge for 10 minutes at 3000rpm and [math]\displaystyle{ 4^o }[/math]C.
- Remove supernatant and replace with 10% original volume TSS.
- Make 100uL aliquots and store at [math]\displaystyle{ -80^o }[/math]C.
TSS (50 mL)
- 5g PEG 8000
- 1.5 mL 1M MgCl2
- 2.5 mL DMSO
- LB to 50 mL
