title: RNA-Seq of mus musculus: adult male liver
design: Liver samples of three LC, PPC, HFD and HFD+PPC mice were detected. For mRNA library construction and deep sequencing, RNA samples were prepared by using the TruSeq RNA Sample Preparation Kit according to the manufactures protocol. Briefly, the poly-A containing mRNA molecules were purified from the 3 g of total RNA by using poly-T oligo-attached magnetic beads. The cleaved RNA fragments were reversely transcribed into first strand cDNA using random hexamers, following by second strand cDNA synthesis using DNA Polymerase I and RNase H. The cDNA fragments were purified, end blunted, 'A' tailed, and adaptor ligated. PCR was used to selectively enrich those DNA fragments that have adapter molecules on both ends and to amplify the amount of DNA in the library. The number of PCR cycles was minimized to avoid skewing the representation of the library. The library was qualified by Agilent 2100 bioanalyzer and quantified by Qubit and qPCR. The produced libraries were sequenced on on the HiSeq 2505 platform.