title: GSM6368717: Bulk RNA-seq Ngn2 rep2; Mus musculus; RNA-Seq
library_construction_protocol: The cortex of post-natal mice aged P5/6 was isolated, mechanically dissociated to obtain a single cell suspension and cultured in medium containing 10% FBS, glucose, pen/strep, B27 supplement, epidermal growth factor and fibroblast growth factor. After culturing for 7 days, the astrocyte enriched culture was reseded on to poly-D-lysine coated 6 well plates and infected with a doxycycline-inducible lentiviral construct. The lentiviral constructs were induced by addition of doxycycline (2ug/ml) for 2 consecutive days in medium containing glucose, pen/strep, glutamine and B27 supplement.  At 48 hours post-doxycycline induction, the transduced astrocytes were isolated with FACS  based on GFP expression Directly after FACS isolation, cell were resuspended in TRIzol, the RNA was extracted using RNA Clean and Concemtrator kit from Zymo Research (cat No. R1013) and the RNA-seq libraries were prepared using NEBNext Ultra II FS DNA library Prep kit from Illumina (#E7805S) according to manufacturer's instructions