GSM7211185: Multiplexed scRNA, 5 individuals-2; Homo sapiens; RNA-Seq
PBMCs were thawed and washed with PBS (no calcium or magnesium), and the pellet resuspended in 100µL of cell staining buffer. The cells were incubated with Fc receptor blocking solution (5 µL, TruStain). Cells were incubated on ice for 10 minutes. A Total Seq C master mix containing surface antibodies (CD3 clone UCHT1 #300479, CD4 clone SK3 #344651, CD8a clone SK1 #344753, CD14 clone 63D3 #367137, CD16 clone 3G8 #302065, CD69 clone FN50 #310951) was prepared. 3µL of the master mix was added each sample, which were incubated on ice for 30 minutes. The cells were washed three times and resuspended in 0.04% BSA in PBS. We submitted twenty thousand cells that were loaded onto the Chromium Single Cell 5' ssay (10x Genomics). Cells were encapsulated and feature barcoded, with library preparation, cDNA amplification and sequencing (50bp paired-end reads, on NovaSeq 6000 S2 platform).