RNA-Seq of pig blood: Large White Restricted Feeding 21 week
High-quality RNA (RIN > 7.5) was used for the preparation of cDNA libraries according to Illuminas protocols (Illumina TruSeq RNA sample prep kit for mRNA analysis). Briefly, poly-A mRNA was purified from 4g of total RNA, fragmented and randomly primed for reverse transcription to generate double stranded cDNA. The cDNA fragments were then subjected to an end repair process, consisting of the addition of a single A base, and the ligation of indexed Illumina adapters at both ends of cDNA. These products were then purified and enriched by PCR to create the final bar-coded cDNA library. After quality control and quantification, cDNA libraries were sequenced on 2 lanes on the NovaSeq6000 S4 (Illumina NEB, USA).