title: GSM6670925: P217; Homo sapiens; RNA-Seq
library_construction_protocol: Venous blood samples (4 mL) were collected from each participant into a BD Vacutainer EDTA tube (BD, Cat#:367861). The Ficoll-Paque medium was used for density gradient centrifugation to obtain peripheral blood mononuclear cells (PBMCs). After trypan blue staining, cells in each sample for the construction of a single-cell RNA library should exceed the cell viability of 80%. Cell suspension with the concentration of 700-1200 cells/μL was prepared for 10x Genomics ChromiumTM. The single-cell library was conducted using Chromium Next GEM Automated Single Cell 3ʹ Library and Gel Bead Kit v3.1 (10×Genomics, Cat#: PN-1000141) according to the manufacturer's protocol. Briefly, cell suspension, enzymes and partitioning oil loaded onto Chromium Next GEM Automated Chip G (10×Genomics, Cat#: PN-1000136) to generate single-cell gel beads in emulsion (GEMs). Barcoded cDNA was generated from poly-adenylated mRNA of captured cells in GEMs. cDNA was PCR-amplified prior to library construction. The constructed libraries were sequenced with 150-bp paired-end reads on an Illumina NovaSeq 6000 platform.