ATCC medium: 803 M7 medium
L-Methionine solution (1.5 g/L)............... 30.0 ml
Glucose solution (270 g/L).................... 20.0 ml
Buffer Solution (see below)................... 20.0 ml
Yeast extract solution (25 g/L).............. 200.0 ml
Dialyzed Fetal Bovine Serum (see below)...... 100.0 ml
Distilled water.............................. 630.0 ml
Filter-sterilize the glucose and L-methionine solutions. Process fetal calf serum as described below. Other ingredients may be autoclaved at 121C for 15 minutes.
Combine ingredients aseptically.
Solution :
Buffer......................18.1 g
KH2PO4 Na2HPO4 ..................... 25.0 g
Distilled water.............. 1.0 L
Dialyzed Fetal Bovine Serum: Dialyze the heat-inactivated serum at 0-4C against 10 volumes of water. Clean the dialysis tubing before use by boiling in a solution of 0.37 g/L of EDTA and rinsing with water. Change the water four times at 8-16 hour intervals. Centrifuge the dialyzed serum for 30 minutes at 35,000g and filter-sterilize.