213. PYG MEDIUM (modified)
  • Tripticase peptone 5.00 g
  • Peptone 5.00 g
  • Yeast extract 10.00 g
  • Beef extract 5.00 g
  • Glucose 5.00 g
  • K2HPO4 2.00 g
  • Tween 80 1.00 ml
  • Resazurin 1.00 mg
  • Salt solution (see below) 40.00 ml
  • Distilled water 950 ml
  • Haemin solution (see below) 10.00 ml
  • Vitamin K1 solution (see below) 0.20 ml
  • Cysteine-HCl x H2O 0.50 g

 

The vitamin K1, haemin solution and cysteine are added after the medium has been boiled and cooled under N2. Adjust pH to 7.2 using 8N NaOH. Distribute under N2 and autoclave.

For solid medium add 15 g of agar powder. 

Salt solution:
  • CaCl2 x 2H2O 0.25 g
  • MgSO4 x 7H2O 0.50 g
  • K2HPO4 1.00 g
  • KH2PO4 1.00 g
  • NaHCO3 10.00 g
  • NaCl 2.00 g
  • Distilled water 1L
Haemin solution:
50 mg haemin dissolved in 1ml 1N NaOH.
Make up to 100 ml with distilled water.
Store refrigerated.

Vitamin K1 solution:
Dissolve 0.10 ml of vitamin K1 in 20 ml 95% ethanol and filter sterilize.
Store refrigerated in a brown bottle.

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