GSM3332256
GSM3332256: Ctrl-P10-3; Mus musculus; RNA-Seq
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.
GEO
GSM3332256
GEO Accession
GSM3332256