GSM1893650: ND.rc; Homo sapiens; RNA-Seq
Total RNA was extracted using the ZR RNA MiniPrep RNA Isolation Kit (Zymo Research), and treated with RNase-free DNase I (Thermo Scientific). cDNA libraries were constructed following TruSeq Stranded mRNA Library Prep Kit (Illumina, San Diego, CA) for next-generation sequencing. Briefly, 400 ng total RNA was used as starting material and PolyA-selected mRNAs were purified, size-fractioned, and subsequently transcribed into single-stranded cDNA by random hexamer priming. Following second-strand synthesis, double-stranded cDNAs were blunt-end fragmented and indexed using adapter ligation, after which they were amplified and sequenced according to manufacturer's protocol (Illumina).