Gibco™ DMEM/F-12, GlutaMAX™ supplement

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	DMEM/F12-GLUTAMAX-type medium containing 4% B27 supplement without vitamin A (Invitrogen), 1% N2 supplement (Invitrogen), and 55 M 2-mercaptoethanol (2-ME). To induce forebrain fate, the medium was supplemented with 5µM Y-27632 and 200ng/ml recombinant mouse Noggin (R&D Systems, 1967-NG-025)	After 2 days, EBs were collected and plated onto 10-cm bacterial Petri dishes in 10 ml neuronal medium as above with the addition of B27 supplemented with vitamin A.

Protocol tips
DMEM/F12-GLUTAMAX-type medium containing 4% B27 supplement without vitamin A (Invitrogen), 1% N2 supplement (Invitrogen), and 55 M 2-mercaptoethanol (2-ME). To induce forebrain fate, the medium was supplemented with 5µM Y-27632 and 200ng/ml recombinant mouse Noggin (R&D Systems, 1967-NG-025)
Downstream tips
After 2 days, EBs were collected and plated onto 10-cm bacterial Petri dishes in 10 ml neuronal medium as above with the addition of B27 supplemented with vitamin A.

Publication protocol

DMEM/F12-GLUTAMAX-type medium containing 4% B27 supplement without vitamin A (Invitrogen), 1% N2 supplement (Invitrogen), and 55 M 2-mercaptoethanol (2-ME). To induce forebrain fate, the medium was supplemented with 5µM Y-27632 and 200ng/ml recombinant mouse Noggin (R&D Systems, 1967-NG-025)

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Manufacturer protocol

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