title: GSM6208554: WKD-Paracancerous; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was extracted and purified using mirVana™ miRNA Isolation Kit (Cat#AM1561, Ambion, Austin, TX, US) following the manufacturer's instructions and checked for a RIN number to inspect RNA integration by an Agilent Bioanalyzer 2100 (Agilent technologies, Santa Clara, CA, US). Paired-end libraries were synthesized by using the mRNA-seq Lib Prep Kit for Illumina  (ABclonal, China) following Sample Preparation Guide. Briefly, The poly-A containing mRNA   molecules were purified using poly-T oligo-attached magnetic beads.Following purification, the   mRNA is fragmented into small pieces using divalent cations under 94℃ for 8 min. The cleaved   RNA fragments are copied into first strand cDNA using reverse transcriptase and random primers.   This is followed by second strand cDNA synthesis using DNA Polymerase I and RNase H. These   cDNA fragments then go through an end repair process, the addition of a single 'A' base, and then   ligation of the adapters. The products are then purified and enriched with PCR to create the final   cDNA library