title: GSM3332252: Ctrl-P6-2; Mus musculus; RNA-Seq
library_construction_protocol: Control (ERRαHet/ERRγWT) and KO (ERRαKO/ERRγKO) mouse hearts were rapidly dissected on ice 10am – 12pm of the day.  Ventricles were freshly processed or flash frozen in liquid nitrogen and subsequently kept at -80°C before nuclear isolation. Tissues were dounced 15 times with loose pestle and then 10 times with tight pestle. For nuclei purification, samples were ultracentrifuged at 25,000 rpm at 4 °C for 2 hours. Single nucleus RNA sequencing was performed using the Drop-Seq method. Briefly, single nuclei, polyA-containing barcoded beads (ChemGenes), and droplet generation oil (Bio-Rad) were each flowed into individual input ports on a microfluidic device (uFluidix). Nuclei in droplets were lysed and hybridized to beads. Beads were isolated and subjected to reverse transcription (Thermo Fisher) and exonuclease I (NEB) treatment, followed by PCR of full length transcripts (Kapa). Final libraries were created by tagmentation of cDNA (Illumina), and amplified by PCR and purified by SPRI beads.
attributes.tag: GEO Accession
attributes.value: GSM3332252