title: GSM6368745: Yy1_scRNAseq_cell_multiplex_library; 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 5 days, the astrocytes were infected with a lentivirus which constitutively expressed the Cre protein and tdTomato fluorescent protein. 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, which expressed Ngn2. The Ngn2 expressing lentiviral construct was induced by addition of doxycycline (2ug/ml) for 2 consecutive days in medium containing glucose, pen/strep, glutamine and B27 supplement The experimental conditions were labelled according to manufacturer's instructions with the following CellPlex reagents from the 3' CellPlex Kit set A (10x genomics, PN: 1000261): Yy1_WT (CMO309), Yy1_KO (CMO310), Yy1_WT/Ngn2+ (CMO311), Yy1_KO/Ngn2+ (CMO312). Approximately, 25,000 events per condition were FACS sorted (Yy1_WT; untransduced, Yy1_KO; RFP+, Yy1_WT/Ngn2+; GFP+, Yy1_KO/Ngn2+; RFP+GFP+)  into an eppendorf tube. Approximately, 33,000 cells were loaded onto a Chromium Next GEM ChIP G (10x Genomics, PN:2000177) to obtain a targeted cell recovery of 20,000 cells. The gene expression library (PN:3000431) and the cell multiplexing library (PN:3000482) were prepared according to manufacturer's protocol (CG000388 Rev A). The gene expression library and the cell multiplexing library were quality controlled using the Agilent 2100 Bioanalyser (Agilent) and the libraries were sequenced according to manufacturer's specifications