HG Agar

• Begin with approx. 700 mLs of DI H₂O.
• Weigh out and add ingredients with constant stirring:

• NaCl 3g
• Bacto Peptone 20g
• Bacto Agar 30g

• Bring up to 970 mL with DI H₂O
• Autoclave
• Allow to cool in 50˚C water bath
• Add:

• 1 M CaCl₂ 1mL (final conc. 1mM)
• 1 M MgSO₄ 1mL (final conc. 1mM)
• 1 M KH₂PO₄ (pH 6.0) 25mL (final conc. 25mM)
• 5mg/mL cholesterol

in 95-100% ethanol 4mL

Protocol from Coleen T. Murphy