Lidstrom:Preparing Freezer Cell Stocks: Difference between revisions
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*Grow ~ 5 mL of E. Coli overnight with appropriate antibiotics. | *Grow ~ 5 mL of E. Coli overnight with appropriate antibiotics. | ||
*Centrifuge for 5-10 minutes & pour off supernatant. | *Centrifuge for 5-10 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 | *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. | ||
*Test stock after frozen a while. | *Test stock after frozen a while. | ||
Revision as of 18:29, 10 February 2012
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DMSO
- Grow ~ 5 mL of E. Coli overnight with appropriate antibiotics.
- Centrifuge for 5-10 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.
