Gibco™KnockOut™ DMEM

Protocol tips

Upstream tips	Protocol tips	Downstream tips
∼5 × 104 undifferentiated hESCs (∼50% confluency in a six-well plate) were seeded onto one well of a six-well plate coated with Matrigel.	differentiation medium (KODMEM supplemented with 10% FBS, 1 mM l-glutamine, 0.1 mM nonessential amino acids, 50 ng ml−1 BMP4 and BMP8a (R&D systems)) for 1 h at 37 °C with 5% CO2.	At day 7, the differentiated hESCs were incubated with differentiation medium containing 0.35 ng ml−1 GDF9:BMP15 heterodimer (prepared as described previously in Matzuk's lab39) and 10 ng ml−1 EGF (R&D) for 1 day, followed by new differentiation medium containing 50 ng ml−1 GDF9 and 25 ng ml−1 BMP15 (R&D systems)

Upstream tips
∼5 × 104 undifferentiated hESCs (∼50% confluency in a six-well plate) were seeded onto one well of a six-well plate coated with Matrigel.
Protocol tips
differentiation medium (KODMEM supplemented with 10% FBS, 1 mM l-glutamine, 0.1 mM nonessential amino acids, 50 ng ml−1 BMP4 and BMP8a (R&D systems)) for 1 h at 37 °C with 5% CO2.
Downstream tips
At day 7, the differentiated hESCs were incubated with differentiation medium containing 0.35 ng ml−1 GDF9:BMP15 heterodimer (prepared as described previously in Matzuk's lab39) and 10 ng ml−1 EGF (R&D) for 1 day, followed by new differentiation medium containing 50 ng ml−1 GDF9 and 25 ng ml−1 BMP15 (R&D systems)

Publication protocol

differentiation medium (KODMEM supplemented with 10% FBS, 1 mM l-glutamine, 0.1 mM nonessential amino acids, 50 ng ml−1 BMP4 and BMP8a (R&D systems)) for 1 h at 37 °C with 5% CO2.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for Gibco™KnockOut™ DMEM below.

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