Lidstrom:Preparing Freezer Cell Stocks

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(New page: Back to protocols Grow ~ 5 mL of E. Coli overnight with appropriate antibiotics. Centrifuge for 5-10 minutes & pour off supernatant. Resuspend in 900 uL of HY (hyp...)
(DMSO)
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Back to [[Lidstrom: Protocols|protocols]]
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Back to [[Lidstrom:Protocols|protocols]]
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Grow ~ 5 mL of E. Coli overnight with appropriate antibiotics.
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==DMSO==
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Centrifuge for 5-10 minutes & pour off supernatant.
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*Grow ~ 5 mL of E. Coli overnight with appropriate antibiotics.
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Resuspend in 900 uL of HY (hypho) and 100 uL of filter-sterilized DMSO.
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*Centrifuge for ~2+ minutes & pour off supernatant.
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Store at -80oC.
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*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.
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Test stock after frozen a while.
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*Store at -80oC.
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*Test stock after frozen a while.
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==Glycerol==
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*

Revision as of 17:08, 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.

Glycerol

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