培養(yǎng)基配方
The difference between this medium and "normal" SP4 medium (Medium 1076) is that the yeast extract was left out and DNA was added instead. Since most mycoplasmas use either glucose or arginine as their energy source one is added to the media. Do not combine both in one medium, arginine can inhibit the growth of some glucose utilizers (unknown reason). For 200 ml of medium
PPLO broth 0.70 g
Tryptone 2.00 g
Bacto-Peptone (BD) 1.00 g
distilled water 130.00 ml
DNA (fish sperm; SERVA) 0.04 g
Adjust pH to 7.4; autoclave at 121?C for 15 minutes.
Add aseptically:
CMRL-1066 medium (10x concentrated; GIBCO) 10 ml
Yeastolate (BD; 2%, autoclaved) 20 ml
Fetal bovine serum (heat-inactivated) 17 ml
Swine serum (heat-inactivated) 17 ml
As energy source add either
Glucose (50% aqueous solution) 2 ml
or
Arginine HCl (50% aqueous solution) 2 ml
For Ureaplasma add instead of Glucose or Arginine Urea 0.4 g
For agar plates use only 126 ml of distilled water and add 2 g of agar. Medium containing agar should be placed at 55?C before adding supplements.
