GSM6647633: P B-cell; Homo sapiens; RNA-Seq
CD19+ B cells were sorted from fresh isolated PBMC using MoFlo XDP (Beckman) and resuspended with PBS supplied with 2%BSA CD19+ B cells were processed using the 10x Genomic Chromium Single Cell Platform at a concentration of 700-1,000 cells per microliter, as described in the manufacturer's protocol. Approximately 8,000-10,000 cells were loaded onto 30 library chips following the Single Cell 30 Reagent Kits (v2) User Guide. The cells were then partitioned into GelBeads in Emulsion in the GemCode instrument, where cell lysis and barcoded reverse transcription of RNA occurred, followed by amplification, enzymatic fragmentation, 50 adaptor, and sample index attachment. The libraries were constructed using Chromium Single Cell 30 Reagent Kits (v2): Single Cell 30 Library & Gel Bead Kit v2 (PN-120237), Single Cell 30 Chip Kit v2 (PN-120236), and i7 Multiplex Kit (PN-120262) (10x Genomics).