GSM9411141: SC06_sFGR_Larger; Homo sapiens; RNA-Seq
Dissociated cells were collected at 20 min intervals to maximize yield and viability. Single-cell suspensions were sequentially filtered through a 40 μm nylon strainer, treated with 1× Red Blood Cell Lysis Solution (Thermo Fisher Scientific, Cat. no. 00-4333-57), and washed with DPBS containing 2% FBS. Single-cell suspensions were processed using the Chromium Single Cell 3′ v3 platform (10x Genomics) following the manufacturer's protocols (CG000204 Rev B). Briefly, individual cells were partitioned into Gel Bead-In-Emulsions (GEMs) containing barcoded oligonucleotides with unique molecular identifiers (UMIs). After cell lysis and reverse transcription, cDNA molecules were tagged with cell-specific barcodes and UMIs. Libraries were generated through second-strand synthesis, adaptor ligation, and amplification, enriching for 3′ transcript ends. Quality and concentration of libraries were assessed using the High Sensitivity DNA Kit (Agilent Bioanalyzer 2100) and Qubit High Sensitivity DNA Assay Kit (Thermo Fisher Scientific). Libraries were sequenced on an Illumina NovaSeq 6000 platform using paired-end 150 bp reads. Single-cell libraries were prepared using the Chromium Single Cell 3′ v3 platform (10x Genomics) according to the manufacturer's protocol