RNA-Seq of human: adult heart: Sample heart_1_rna
The cytoplasmatic rRNA removal was performed for each total RNA sample using the Ribo-Zero rRNA removal Kit (Epicentre, Madison, WI, USA). The rRNA-depleted RNA was used to prepare the stranded-oriented RNA-seq library using the TruSeq Stranded Total RNA Sample Prep Kit (Illumina, San Diego, CA, USA), according to the manufacturer’s instructions. Briefly, each RNA was chemically fragmented prior to the random priming reverse transcription reaction for first strand cDNA generation. The fragmentation step resulted in a RNA-seq library including inserts ranging in size from approximately 100-400 bp. During the second strand synthesis, dUTP was incorporated in place of dTTP, thus preventing amplification of this strand during the subsequent PCR step and retaining strand information. cDNA libraries were sequenced on the Illumina HiSeq2500 platform. Paired-end reads of 100 nt were generated for each fragment.