title: GSM6368729: Bulk ATAC-seq PmutNgn2 rep2; Mus musculus; OTHER
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 of 50,000 cells were transposed with a modified Tn5 transposase (Illumina Tagment DNA TDE1 Enzyme and Buffer kit, 20034198) according to the OMNI-ATAC-seq protocol (Corces et al., 2017), the trasposed DNA was isolated using the DNA clean and concentrator kit from Zymo Research (D4014) and the ATAC-seql ibraries were prepared using NEBNext Ultra II  Q5 Master mix  from NEB (M0544S) according to manufacturer's instructions. The size distribution of the ATAC-seq libraries was analysed using an Agilent 2100 Bioanalyser