GSM4007698: TD rep1; Homo sapiens; RNA-Seq
RNA was extracted with Trizol reagent, followed by clean-up and DNase I treatment with QIAGEN RNeasy mini kit in accordance with the prescribed protocol provided with the kit. Quality control was performed with Agilent Bioanalyser. For library construction, we used the TruSeq Stranded mRNA Library Prep Kit (Illumina, San Diego, CA). Briefly, the steps of strand-specific protocol are: first strand cDNA synthesis; second strand synthesized using dUTPs instead of dTTPs; end repair, A-tailing, and adaptor ligation; PCR amplification. Then, each library was diluted to 8 pM for 76 cycles of paired-read sequencing (2 X 75bp) on the Illumina NextSeq 500 per the manufacturer's recommended protocol.