Lidstrom:Preparing Freezer Cell Stocks

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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.
+
*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 21:29, 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.

Glycerol

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