GSM9685694: LV SCI 1d rep1; Rattus norvegicus; RNA-Seq
Frozen heart tissues were homogenized for 20s at 5000 revolutions per minute on the Precellys Evolution homogenizer (Bertin Technologies SAS, Montigny-le-Bretonneux, France) using the 2 mL soft tissue homogenizing ceramic beads CK14 (Bertin Technologies SAS, Cat No. P000912-LYSK0-A). Total RNA was isolated from the extracts using the Maxwell RSC simplyRNA Tissue Kit (Promega Cat. No. AS1340) on the Maxwell RSC instrument (Promega) according to the manufacturer's instructions. Extracted RNA was treated with DNase1 and its quality was analyzed using the High Sensitivity RNA Assay (Agilent Technologies, Cat No. 5067-5579 and 5067-5580) on the TapeStation 4200 (Agilent Technologies, Cat. No. G2991BA). RNA concentrations are measured using the Qubit RNA HS assay (ThermoFisher; Cat No. Q32852) on the Qubit 4 Fluorometer (ThermoFisher).  RNA quality metrics for RNA include the RNA integrity number (RIN) and DV200. Samples meeting pre-specified quality thresholds were advanced to library preparation. 100ng of total RNA was used as input for ribosomal RNA depletion using the DNBSEQ rRNA Depletion kit (Complete Genomics, Cat No. 940-001811-00).  Strand-specific libraries were prepared using the DNBSEQ Fast RNA Library Prep kit (Complete Genomics, Cat No. 940-000963-00). Library quality and size were assessed with the TapeStation 4200 (Agilent Technologies; D1000 Assay Cat No. 5067-5582), and libraries were quantified using the Qubit dsDNA HS Assay (Thermo Fisher; Cat No. Q32854). Libraries were sequenced on a DNBSEQ-G400RS sequencer using paired-end 100 bp reads (PE100).