Difference between revisions of "Lidstrom:Preparing Freezer Cell Stocks"

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(DMSO)
(DMSO)
Line 5: Line 5:
 
*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 20:10, 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.

Glycerol