Team:Cambridge/Protocols/SporeMedia

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Revision as of 13:52, 19 September 2012

Chemically Defined Sporulation Media (CDSM)

ref.: W. Nicholson & P. Setlow, in Molecular Biological Methods for Bacillus, eds. C. Harwood & S. Cutting, New York: John Wiley, section 9, 1990.


CDSM

Notes: 120mL CDSM makes one batch of spores Adujst stock solutions for each below to pH7.0 with KOH (except for MT mix). Autoclave stock solutions, except MOPS and L-Glutamic acid which are filter sterilised. L-Glutamic acid is made up in 1M HCl

  • 40mM morpholinopropanesulfonic acid (MOPS)
  • 4.0mM KH2PO4
  • 9.5mM (NH4)2SO4
  • 5.0mM L-lactic acid
  • 8.0mM L-Glutamic acid
  • 20mM - D-Glucose
  • 1xMT mix

50xMT mix (trace metal mix)

Note: Add FeCl3 last to avoid precipitation This solutions makes 50L CDSM

For 1L:

  • 2ml 1M HCl
  • 40.6g Mg Cl2
  • 10.29g CaCl2
  • 0.99g MnCl2
  • 13.6g ZnCl2
  • 67.5mg Thiamine-HCL
  • 135mg FeCl3


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