ES cell medium

Reagent	Final concentration	Amount
G-MEM with L-Glutamine and Phenol Red	n/a	153.63 mL
KnockOut Serum Replacement (KSR)	20%	40 mL
Sodium pyruvate (100 mM)	1 mM	2 mL
2-mercaptoethanol (2-ME)	0.1 mM	2 mL
Non-essential amino acids (NEAA)	1/100 of the stock	2 mL
Recombinant mouse leukemia inhibitory factor (LIF)	1 ng/mL	20 μL
Stemolecule CHIR99021 (3 mM stock)	3 μM	200 μL
Stemolecule PD0325901 (2 mM stock)	0.5 μM	50 μL
Penicillin/Streptomycin stock	20 U/mL/20 μg/mL	100 μL
Total	n/a	200 mL

Store at 4°C for 3 weeks.