Chromium single cell 3 reagent kits v2 and v3

The scRNA-Seq samples (freshly isolated and MACS-sorted CBMCs and freshly isolated and MACS-sorted HUVECs from the same individual human) were applied to a single cell master mix with lysis buffer and reverse transcription reagents, following the Chromium Single Cell 3’ Reagent Kits V2 and V3 User Guide (10x Genomics). This was followed by cDNA synthesis and library preparation. All libraries were sequenced in Illumina NovaSeq6000 platform in paired-end mode (28 bp + 91 bp). The total number of CBMCs and HUVECs were 4,477 cells and 13,651 cells, respectively. Next, 85,000 reads per cell were generated and 94% of the sequencing reads reached a q score of at least 30 (Q30) in CBMC, while 28K reads per cell were generated and 95% of the sequencing reads reached Q30 in HUVECs.

In total, the cell viability of all samples is above 90%, 10,000 cells per sample were loaded into a Chromium Single-Cell 3′ Chip Kit v2 and processed using a Chromium single cell 3′ Reagent Kits (v2 and v3) (10× Genomics) according to the manufacturer’s instructions. Libraries were constructed using the Single 3′ Library and Gel Bead Kit v2 (PN-120237) and Chromium i7 Multiplex Kit v2 (PN-120236).