ATCC Medium: 2764 SP4-Z MEDIUM
PPLO Broth………………………3.5 g
Tryptone………………………….10.0 g
Peptone…………………………...5.0 g
DI Water……………………….….640.0 ml
DNA (fish sperm)………………...0.2 g (spectrum DE120; cas# 100403-24-5)
Adjust pH to 7.4 and autoclave at 121?C.
Add aseptically:
CMRL-1066 medium (10x concentrated;ATCC 20-2207)…….50 ml
Yeastolate (2%, autoclaved)…………………………….100 ml
Fetal bovine serum (heat-inactivated)…………………85 ml
Swine serum (heat-inactivated)…………………………85 ml
Glucose (50% aqueous solution)……………………….20 ml
L-cysteine solution (see below)…………………………..10 ml
NAD solution (see below)…..………………………….…10 ml
Medium containing agar should be placed at 55?C before adding supplements.
Re-adjust pH to 7.4 after addition of supplements
L-cysteine solution
L-cysteine………………………………………………….0.01 g
DI Water……………………………………………………10 ml
NAD solution
NAD…………………………………………………………0.1 g
DI Water…………………………………………………….10 ml
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