ATCC Medium 798 (see below) with fresh yeast extract replaced by water
ATCC Medium 798:
1. Autoclavable Fraction:
D-Glucose......................................0.1 g
D-Fructose.....................................0.1 g
Sucrose........................................1.0 g
Tryptone (BD 211705)...........................1.0 g
Sorbitol.......................................7.0 g
PPLO Broth w/o CV (BD 255420)..................2.1 g
Peptone........................................0.8 g
Distilled water...............................70.0 ml
Adjust pH to 7.8 with N NaOH. Autoclave 15 minutes at 121C. Cool to room temperature.
2. Aseptically add the following nonautoclavable components:
Schneider's Drosophila Medium (GIBCO 11720)..160.0 ml
Fetal bovine serum (heat-inactivated).........50.0 ml
Yeast Extract Solution (GIBCO 18180)..........10.0 ml
Phenol red (0.5%)..............................1.2 ml
Final pH of complete medium should be approximately 7.4.
Preparation of M-1 Solid Medium:
1. Add 5.0 g Agar, Noble (BD 214230) to autoclavable fraction. Autoclave 15 minutes at 121C. Equilibrate at 56C.
2. Equilibrate heat-inactivated serum, phenol red, and yeast extract at 56C.
3. Equilibrate Schneider's rapidly to 37C.
4. Mix components rapidly. Pour immediately.