title: GSM4585218: TA00_16; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was extracted from whole blood samples using TRIzol (Bio Basic Inc.) according to the manufacturer's instructions, and analyzed using RNA Nano 6000 Assay Kit of the Bioanalyzer 2100 system (Agilent Technologies, CA, USA) and Qubit® RNA Assay Kit in Qubit® 2.0 Flurometer (Life Technologies, CA, USA). mRNA sequencing libraries were built using 3 µg RNA and NEBNext® UltraTM RNA Library Prep Kit for Illumina® (NEB, USA) following the manufacturer's recommendations. Poly-T oligo-attached magnetic beads were employed to isolate poly-A RNA. First strand cDNA was synthesized using random hexamer primer and M-MuLV Reverse Transcriptase (RNase H-). Second strand cDNA synthesis was subsequently performed using DNA Polymerase I and RNase H. After adenylation of 3'ends of DNA fragments, NEBNext Adaptor with hairpin loop structure were ligated to prepare for hybridization. At last, PCR products were purified (AMPure XP system) and library quality was assessed on the Agilent Bioanalyzer 2100 system. The clustering of the index-coded samples was performed on a cBotCluster Generation System using TruSeq PE Cluster Kit v3-cBot-HS (Illumia) according to the manufacturer's instructions. After cluster generation, the library preparations were sequenced on an Illumina Hiseq platform and 125 bp/150 bp paired-end reads were generated.