GSM1807988: Liver_16; Homo sapiens; RNA-Seq
In brief, 0.5-1.0 µg of total RNA was twice selected for mRNA by oligo (dT) and then fragmented by heating. First strand cDNA was synthesized using Superscript III reverse transcriptase and random hexamer primers. After second strand synthesis by DNA polymerase I and with dUTP in place of dTTP, double stranded cDNA was end-repaired and A-tailed prior to ligation of Illumina adaptors including DNA indices. Libraries were made strand-specific by digestion with Uracil-DNA Glycosylase prior to PCR amplification. Bead-based clean-up was incorporated after each enzymatic reaction and libraries were checked by flash gel and Bioanalyzer analysis. Total RNA was isolated from tissues using TRIzol Reagent.