GSM5766276: scRNA_Tcf7flox_1; Mus musculus; RNA-Seq
cell isolation protocolCells were isolated from bone marrows. Isolated cells were further sorted based on their surface markers. The scRNA-seq libraries were generated using Chromium Single Cell 3' Library & Gel Bead Kit v3 (10X Genomic) according to manufacturer's protocol. Briefly, 15,000 live cells were FACS-sorted and used to generate single-cell gel-bead in emulsion (GEM). After reverse transcription, GEMs were disrupted. Barcoded cDNA was isolated and amplified by PCR (12 cycles). Following fragmentation, end repair, and A-tailing, sample indexes were added during index PCR (8 cycles). Indexed libraries were multiplexed and sequenced on an Illumina NOVO Seq instruments according to the manufacturer's instructions (26 cycles of Read 1, 8 cycles of i7 Index, and 98 cycles of Read2).