GSM8775449: Vascular smooth muscle cells, AAA, DM; Homo sapiens; RNA-Seq
Cell cultures for the 10x Genomics Flex assay were trypsinized, washed in PBS and resuspended in fixation buffer according to the 10x Genomics protocol for cell/nuclei fixation (CG000478). Fixed samples were stored at -80C and shipped to the Genomics Shared Resource at Dartmouth Cancer Center. Cell suspensions were counted on a Cellometer K2 automated cell counter (Revity) using AO/PI fluorescent live/dead staining. Up to 1x106 cells were used as input into the hybridization reaction using the Human Transcriptome Probe Kit which contains sample specific barcodes. Four barcoded samples were pooled together in equal amounts and loaded onto a Chromium X instrument targeting 10,000 cells per sample. Libraries were prepared according to the manufacturer's protocol (CG000527) and sequenced on a NextSeq2000 instrument (Illumina) targeting 25,000 reads/cell.