title: GSM9075923: PBMC_Placebo_pooled; Homo sapiens; RNA-Seq
library_name: GSM9075923
library_construction_protocol: PBMC-derived cells were thawed in 10% FBS/DMEM at 37°C, washed twice with cold calcium/magnesium-free 0.04% BSA/PBS, and resuspended in cold Stain Buffer (BD Biosciences). Cell viability and concentration were assessed using a LUNA-FX7™ cell counter (Logos Biosystems) with AO/PI staining. Each sample was labeled with antibody-conjugated DNA barcodes and incubated for 20 min at room temperature. Cells were washed three times with Stain Buffer and resuspended in cold Sample Buffer (BD Biosciences) before pooling and recounting. Single-cell capture was performed using the BD Rhapsody HT Xpress system and 8-lane cartridge (BD Biosciences). Cell lysis, mRNA capture on barcoded beads, cDNA synthesis, and Exonuclease I treatment were conducted per BD protocol using BD cDNA Kit (Cat# 633773). Whole transcriptome libraries were constructed using BD Rhapsody WTA Amplification Kit (Cat# 633801) through RPE and index PCR. Sample tag libraries were generated via nested PCR and indexing.