title: Digital expression analysis of glioma neural stem cells
design: Digital expression analysis of glioma neural stem cells
library_construction_protocol: GNS and NS cells were cultured in N2B27 serum-free medium, a 1:1 mixture of DMEM/F-12 and Neurobasal media (Invitrogen) augmented with N2 (Stem Cell Sciences) and B27 (Gibco) supplements. Self-renewal was supported by the addition of 10 ng/ml EGF and 20 ng/ml FGF-2 to the complete medium. Cells were plated at 20,000/cm2 in laminin-coated vessels (10 ug/ml laminin-1 (Sigma) in PBS for 6-12 h), passaged at confluence using Accutase dissociation reagent (Sigma) and were typically split at 1:3 for NS cells and 1:3 to 1:6 for GNS cells. TRIzol, phenol/chloroform extraction