GSM5531447: PA_hypoxia; Mus musculus; RNA-Seq
The pulmonary arteries were chopped mechanically and placed in 1mg/ml collagenase (Roche) and 3 U/ml elastase (Worthington). Following cellular dissociation, we barcoded unique mRNA molecules of each cell using our 10x Genomics Chromium platform (3' v3.1 kit), a droplet-based microfluidic system, and performed reverse transcription, cDNA amplification, fragmentation, adaptor ligation, and sample index PCR according to the manufacturer's protocol. High sensitivity DNA bioanalyzer traces of cDNA after barcoding and of the final cDNA library were evaluated for quality control. The final cDNA libraries were sequenced on a HiSeq 4000 Illumina platform in our core facility aiming for 150 million reads per library.