RNAseq of mus musculus heart
RNA was first poly(A) selected using poly(T) oligo-attached magnetic beads and mRNA was purified and fragmented using divalent cations under elevated temperature. Random primer reverse transcription was done for RNA fragments before second-strand cDNA synthesis with DNA Polymerase I and RNase H. After end repair and A-tailing, indexing adaptors were ligated. The products were then purified and amplified to create the final cDNA libraries. After library validation and quantification (Agilent tape station), equimolar amounts of library were pooled. The pool was quantified by using the Peqlab KAPA Library Quantification Kit and the Applied Biosystems 7900HT Sequence Detection System.