GSM9699022: B cells, Whole transcriptome + TCR + BCR, patient 3 baseline; Homo sapiens; RNA-Seq
PBMCs were isolated using Ficoll density gradient centrifugation and were cryopreserved at -80C for 24 hours before storage in liquid nitrogen. Cells were rapidly thawed at 37 °C, treated for 15 min with DNase I and MgCl2 in complete RPMI, and washed. PBMCs were labeled with oligonucleotide barcodes (sample tags) using the BD Human Single Cell Sample Multiplexing Kit (Waters Biosciences). B cells were enriched from remaining PBMCs using the EasySep Human Pan-B Cell Enrichment Kit (StemCell Technologies). Single-cell capture and library preparation were performed using the microwell-based BD Rhapsody platform (Waters Biosciences) using the BD RhapsodyTM Enhanced Cartridge Reagent Kit v3. cDNA amplification and library preparation for whole transcriptome analysis (WTA), T cell receptors (TCR), B cell receptors (BCR), and sample tags were performed using BD Rhapsody kits according to manufacturer instructions.