Preparing chemically competent cells
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(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 09:31, 26 May 2005
- Grow a 5-25mL culture to an OD600 of 0.2
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
