Team:Cambridge/Protocols/MediumB

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