Lidstrom:Preparing Freezer Cell Stocks: Difference between revisions
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==DMSO== | ==DMSO== | ||
*Grow ~ 5 mL of E. Coli overnight with appropriate antibiotics. | *Grow ~ 5 mL of E. Coli overnight with appropriate antibiotics. | ||
*Centrifuge for | *Centrifuge for ~2+ minutes & pour off supernatant. | ||
*Resuspend in 900 uL of HY (hypho) and 100 uL of filter-sterilized DMSO. Hypho is better than LB for this; I hear it buffers. | *Resuspend in 900 uL of HY (hypho) and 100 uL of filter-sterilized DMSO. Hypho is better than LB for this; I hear it buffers. | ||
*Store at -80oC. | *Store at -80oC. |
Revision as of 15:08, 1 March 2012
Back to protocols
DMSO
- Grow ~ 5 mL of E. Coli overnight with appropriate antibiotics.
- Centrifuge for ~2+ minutes & pour off supernatant.
- Resuspend in 900 uL of HY (hypho) and 100 uL of filter-sterilized DMSO. Hypho is better than LB for this; I hear it buffers.
- Store at -80oC.
- Test stock after frozen a while.