GSM8640321: PC, donor 4, scRNAseqVDJ; Homo sapiens; RNA-Seq
Bone marrow cells were stained for viability with Zombie-Yellow™ Fixable Viability Kit (1:600) for 20 minutes at room temperature; and afterwards with anti-CD14-BV421, anti-CD16-BV421, anti-CD56-BV421, anti-CD38-APC, anti-CD45-FITC, and anti-CD138-PE/Cy7 (all Biolegend). After the staining, plasma cells (CD45+CD14-CD16-CD56-CD38++CD138++) were sorted on a FACS Aria machine. At the output, 1600-3100 live cells were subjected to 10x Chromium Single Cell sequencing library generation with 10X Chromium 5' GEX/VDJ kits according to the manufacturer's instructions. GEX Library sequencing was performed on an Illumina HiSeq 2500 sequencer to a depth of about 50 million reads each. In all samples, the mean number of reads per cell was greater than 30000. VDJ Libraries were sequenced to a depth of about 30 million reads resulting in 1000 to 10000 read pairs per cell.