GSM9097424: PBMC, healthy, 43F, GEX; Macaca mulatta; RNA-Seq
For each sample, macaques were first sedated with 10-15 mg/kg ketamine or ketamine/dexmedetomidine(0.025mg/kg) intramuscularly. Heparinized blood was collected from the femoral vein. Plasma and peripheral blood mononuclear cells were isolated by whole blood gradient centrifugation on Lympholyte-Mammal (Cedarlane Lab). Plasma was frozen at -20C for subsequent analysis. Single-cell capture and library creation were performed using Chromium single cell 5' reagent kits user guide (v2 chemistry dual index) with feature barcoding technology for cell surface protein and immune receptor mapping according to 10× Genomics specifications. Specifically, PBMCs were first loaded with Single Cell VDJ 5' Gel Beads and partitioning oil onto a Chromium Next GEM Chip K to produce single-cell gel bead-in-emulsions (GEMs). It was followed by gel bead dissolution and cell lysis, allowing for poly-adenylated mRNA transcripts to be reverse-transcribed into 10x-barcoded full-length cDNA. Later, cDNAs were amplified via polymerase chain reaction (PCR) and subsequently sequenced on Illumina Novaseq 6000.One library was first generated to elucidate the mRNA transcript expression (RNA); from which another library was generated to describe specifically B cell receptor (BCR) repertoire (VDJ) for each single B cell.