Truseq sbs and pe cluster v3 kits

RNA was isolated from whole hearts with their atria removed and its quality was assessed using a Bioanalyser (2100 Instrument, Agilent). High-quality RNA (RIN score > 8) was used for library preparation. One microgram of RNA was used to generate sequencing libraries using NEBNext Ultra II Directional RNA library Prep Kit for Illumina (NEB) according to the manufacturer’s recommendation. Purification steps were performed using the ‘ProNex Size-Selective Purification System’ (fragment cut-off size 100 bp). The index primers employed during the library preparation corresponded to the ‘NEBNext Multiplex Oligos for Illumina (Index Primers Set 1)’. Once the libraries were generated, their quality was evaluated on the Bioanalyzer. Those libraries whose electropherograms showed a narrow distribution with a peak size of ∼300 bp were sent for sequencing. Paired-end multiplexed sequencing of libraries to generate reads of 100 bp was performed on a HiSeq 1000 instrument with TruSeq SBS and PE Cluster v3 Kits (Illumina); 70–100 million reads per sample were obtained.
A minimum of four hearts per genotype were used for sequencing and analysis. Data will be made available in the European Molecular Biology Laboratory–European Bioinformatics Institute (EMBL-EBI) database.