title: GSM6368707: Multiome_scRNA-seq_Ngn2; 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, the nuclei were isolated according to 10x Genomic protocol (CG000365) and scRNA-seq libraries (PN-1000230, PN-1000284, PN-3000431) were prepared by using 5000. nuclei for the astrocyte sample and 1250 nuclei for the GFP, Ngn2 and PmutNgn2 samples as input in the 10x Genomics Chromium Next GEM Single Cell mutiome ATAC + Genexpression kit (PN-1000285). The size distribution of the obtained libraries was assessed using an Agilent 2100 Bioanalyser.