GSM9899966: MN5; Homo sapiens; RNA-Seq
Human PBMCs were pre-isolated from EDTA anticoagulated peripheral blood via Ficoll density gradient centrifugation and cryopreserved long-term. Cryopreserved PBMCs were quickly thawed at 37 °C, washed with warm RPMI medium to remove DMSO, then purified with cold PBS + 0.04% BSA, and qualified with >90% cell viability. Library was performed according to the manufacter's instructions (single cell 5' v3 protocol, 10x Genomics).Cell suspension was adjusted to 700–1200 cells/μL and loaded onto 10x Genomics Chromium Next GEM Chip G to generate single-cell GEM droplets. Within each GEM, cell lysis, polyA mRNA capture via 16 bp barcode + 12 bp UMI poly(dT) gel bead oligos, and in-droplet reverse transcription were completed for single-cell RNA extraction, following Lianchuan Biotechnology's 10x scRNA-seq experimental protocol.