GSM5662568: PBS_hashtag; Mus musculus; RNA-Seq
Dissected aortas were processed for digestion for 10 min at 37 °C in an enzymatic mix composed of type II collagenase (10 mg/ml, C6885, Sigma Aldrich) and elastase (1 mg/ml, LS002292, Worthington Biochemistry). Following the digestion, viable cells were enriched using a dead cell removal magnetic kit (130-090-101, Miltenyi Biotech; for non-pooled samples only) and cellular viability, assessed using 0.4% Trypan blue (1450013, Bio-Rad) on an automated cell counter (TC20, Bio-Rad). 15,000 of viable cells per sample (15,000 viable cells per pooled sample and 15,000 viable cells per hashtag in hastagged samples) were loaded on a 10x Genomics Chromium instrument to obtain individual gel beads in emulsion (GEMs). Chromium Single Cell 3′ Library & Gel Bead Kit v2, PN-120237, the Single Cell 3′ Chip Kit v2 PN-120236 and the i7 Multiplex Kit PN-120262, (10x Genomics) were used for library preparation. Illumina HiSeq 4000 as 2 x 150 paired-end reads was used for sequencing (>90% sequencing saturation).