Lidstrom:Preparing Freezer Cell Stocks: Difference between revisions
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(→DMSO) |
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*Centrifuge for ~2+ minutes & pour off supernatant. | *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. | ||
** ''' must use a filter that won't dissolve in DMSO '''. Most of the filters we have are not appropriate. | |||
*Store at -80oC. | *Store at -80oC. | ||
*Test stock after frozen a while. | *Test stock after frozen a while. | ||
Revision as of 21:10, 6 June 2013
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.
- must use a filter that won't dissolve in DMSO . Most of the filters we have are not appropriate.
- Store at -80oC.
- Test stock after frozen a while.
