Team:Cambridge/Protocols/MediumB

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==Medium B==
==Medium B==
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*Autoclave Medium A base before adding glucose, and autoclave Bacillus salts.
*Autoclave Medium A base before adding glucose, and autoclave Bacillus salts.
*Store aliquots of 10X Medium A base 10mL and 10X Bacillus salts 9mL and keep them in the fridge, never use them twice to avoid contamination
*Store aliquots of 10X Medium A base 10mL and 10X Bacillus salts 9mL and keep them in the fridge, never use them twice to avoid contamination
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Revision as of 11:46, 14 September 2012

Medium B

10X Medium A base:

  • Yeast extract 10g
  • Casamino acids 2g
  • Distilled water to 900mL
  • Autoclave then add:
  • 50% glucose, filter sterilized 100mL

10X Bacillus salts:"

  • (NH4)2SO4.3H2O 20g
  • K2HPO4.3H2O 183g
  • KH2PO4 60g
  • Tri-sodium citrate 10g
  • MgSO4.7H2O 2g
  • SDW to 1000mL

Medium A

  • Sterile water 81mL
  • 10X Medium A base 10mL
  • 10X Bacillus salts 9mL
  • L-Tryptophan (11mg/mL) 0.1mL

Medium B

  • Medium A 10mL
  • 50mM CaCl2.2H2O 0.1mL
  • 250nM MgCl2.6H2O 0.1mL

Important

  • Autoclave Medium A base before adding glucose, and autoclave Bacillus salts.
  • Store aliquots of 10X Medium A base 10mL and 10X Bacillus salts 9mL and keep them in the fridge, never use them twice to avoid contamination