nucleic acid sequencing protocol
Libraries were multiplexed in pairs and run on a single lane of the HiSeq 2000 platform (Illumina) to generate 100bp paired-end reads.
nucleic acid extraction protocol
RNA was extracted from 25mg of crushed left ventricular tissue from four BN and four SHR animals without pooling, using Trizol (Invitrogen) according to manufacturer's instructions.
nucleic acid library construction protocol
4ug of total RNA was used to generate RNA-seq libraries using TruSeq RNA kit (Illumina) according to manufacturer's instructions.
Rats were housed in an air-conditioned animal facility and allowed free access to standard laboratory chow and water. All experiments were done in agreement with the Animal Protection Law of the Czech Republic and were approved by the Ethics Committee of the Institute of Physiology, Czech Academy of Sciences, Prague. We collected left ventricular tissue from unfasted males from six week old male BN-Lx/Cub (referred to in this study as BN), SHR/Olalpcv (referred to as SHR) (n = 4 per strain); and from (BNxSHR)F1 and (SHRxBN)F1 (n = 4 per cross); and from male rats from 29 BXH/HXB recombinant inbred strains (n = 2 per strain) (Pravenec et al. 1989) between 9:00am and 10:00am. Left ventricles were snap frozen in liquid nitrogen and stored at -80°C. The BN-Lx/Cub and SHR/Olalpcv lines have been inbred over more than 80 generations (Simonis et al. 2012).