GSM9468283: BBp53n_OPCs; Mus musculus; RNA-Seq
steps to extract nuclei followed a slightly modified version of the 10X nuclei isolation kit (cat# 1000493) protocol for gene expression libraries. The frozen tissue was dissociated in lysis buffer and filtered. At the debris removal step, we replaced the 10X kit solution with Miltenyi's debris removal solution (cat# 130-109-398) and followed Miltenyi's protocol. After debris removal, the nuclei were resuspended in 0.1% BSA in PBS and continued following the 10X nuclei isolation kit protocol. OPC sample derived from cultured cells and followed nuclei isolation protocol withouth debris removal or filtering. Nuclei yield ranged from 5,000-20,000/mg. After nuclei extraction, the purified nuclei were transported to the CUNY ASRC's epigenetics core at a concentration of 10e6/mL for 10X 3' Gene Expression library preparation (Chromium Next GEM Single Cell 3ʹ Kit v3.1, 16 rxns PN-1000268). Samples were sequenced with a target of 10,000 nuclei per sample and 40,000 reads per nuclei a Novaseq 6000 sequencer