ES cell culture medium

Recipe

Reagent	Supplier	Amount to add	Final concentration
Dulbecco’s modified Eagle’s medium (DMEM)	GIBCO/Invitrogen 11960-044	500 ml	N/A
Fetal bovine serum (FBS)	Hyclone SH30070.03	94 ml	15%
10 mM MEM nonessential amino acids	Gibco/Invitrogen 11140-050	6.2 ml	0.1 mM
100 mM sodium pyruvate	GIBCO/Invitrogen 11360-070	6.2 ml	1 mM
55 mM β-mercaptoethanol	GIBCO/Invitrogen 21985-023	620 μl	55 μM
10,000 units/ml penicillin:10,000 units/ml streptomycin:200 mM L-glutamine	GIBCO/Invitrogen 10378-016	6.2 ml	100 units/ml penicillin:100 units/ml streptomycin:2 mM L-glutamine
10⁷ units/ml leukemia inhibitory factor (LIF)	ESGRO, Chemicon ESG1107	62 μl	1000 units/ml