Complete Growth Medium
Culture Medium: Modified Dulbecco's medium (see below), 0.0007%
2-mercaptoethanol, 90%; fetal bovine serum, 10%.
DME w/HEPES (10mM) 100 mL
L-Glutamine (100x) 1 mL
Solution I 1 mL
Non-essential amino acids (100x) 1 mL
NCTC 135 10 mL
Fetal Bovine Serum 12 mL
Solution I (100x)
1. 1320 mg oxalacetic acid (100 mM, MW 132).
2. 80 mg crystalline bovine insulin (20 units/mL, 25 units/mg).
Add 1 and 2, stir at 37°C. Add Na pyruvate 550 mg (50 mM, FW 110). Bring up to
100 mL with distilled water. Stir at 37°C until solution dissolves. Filter,
aliquot and store frozen.
