Lidstrom:Preparing Freezer Cell Stocks

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Revision as of 20:10, 6 June 2013 by Janet B. Matsen (talk | contribs) (DMSO)
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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.

Glycerol