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Human Embryonic Stem Cell (hES) Media

  Small Bottle (~300mL) Large Bottle (~600 mL)
Knockout DMEM 250 mL 500 mL
Serum Replacement (10%) 37.5 mL 75 mL
10mg/ml Pen/Strep 3 mL 6 mL
200 mM Glutamax 3 mL 6 mL
Non-essential amino acids 3 mL 6 mL
B-mercapto ethanol 300 uL 600 uL
Basic FGF ( 20ng/mL) 150 uL 300 uL

- combine all ingredients as indicated
- filter solution through 0.2 uM Nalgene filter unit (250 mL or 500 mL size)

For antibiotic resistance (selection media)

- Puromycin add to a final concentration of 0.8 ug/ml
- Neomycin add to a final concentration of 50 ug/ml
(media does not need to be re-filtered following addition of antibiotics)