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