PPLO Broth w/o CV (BD 255420).......................11.0 g
Tryptone (BD 211705)................................10.0 g
Glucose..............................................5.0 g
Distilled water....................................625.0 ml
Adjust pH to 7.5. Autoclave at 121C for 15 minutes.
Add Aseptically:
Mycoplasma Growth Supplement (CMRL 1066)(10X)*......50.0 ml
Yeast Extract Solution (see below)..................35.0 ml
TC Yeastolate (2% soln.) (BD 255772)...............100.0 ml
Fetal bovine serum (heat-inactivated)**............170.0 ml
Phenol red solution (0.1% soln.)....................20.0 ml
Mycoplasma Growth Supplement:
Prepare by adding 1.0 L of water to one bottle of CMRL-1066 powder. Mix to dissolve and filter-sterilize.
*This supplement is now available as ATCC 20-2206: 98.0 g powder and as ATCC 20-2207: 100.0 ml (10x solution).
**Replaceable with newborn calf serum.
Yeast Extract Solution:
Bakers’ yeast (live, pressed, starch-free).........250.0 g
Distilled deionized water............................1.0 L
Mix yeast and water; autoclave at 121C for 90 minutes at 15 psi pressure.
Allow to stand. Remove supernatant solution. Adjust pH to 6.6- 6.8