N2B27 medium.

Reagent	Amount	Final concentration
DMEM/F12 (Gibco#11330–032)	47.5 mL
Neurobasal media (Gibco#21103–049)	47.5 mL
l-Glutamine (200 mM) (GIBCO#25030)	1 mL	2 mM
β-Nercaptoethanol (10 mM) (Sigma#M7522)	1 mL	0.1 mM
N2 supplement (100 ×) (Invitrogen#17502048)	1 mL	1 ×
B27 supplement (50 ×) (Invitrogen#17504–044)	2 mL	1 ×

Filter and store the medium (final volume 100 mL) at 4°C and use it within 2 weeks. Prepare β-mercaptoethanol 10 mM stock from 14.3 M stock (Sigma-Aldrich M7522) by adding 6.25μL to 9 mL of sterile distilled water.