title: GSM4725674: Mock-3; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was extracted using NucleoSpin® RNA Plus kit (Macherey-nagel) according to the manufacture's instruction, including an on-column genomic DNA digestion step.  Quality and quantity of RNA was checked with Qubit and Bioanalyzer.  Ribosomal RNA (rRNA) depletion was performed using Ribo-Zero Manetic Gold Kit. cDNA libraries were prepared by NEBNext® Ultra II Directional RNA Library Prep Kit.  The purified rRNA-depleted RNA was fragmented to 200 – 500 bp. The fragmented rRNA-depleted RNA was then applied as template to synthesize the first-strand cDNA by using random hexamer-primer and reverse transcriptase. In the second strand cDNA synthesis, the mRNA template was removed, and a replacement strand was generated to form the blunt-end double-stranded (ds) cDNA. The ds cDNA underwent, 3' adenylation and indexed adaptor ligation, followed by treatment with User Enzyme. The adaptor-ligated libraries were enriched and indexed by polymerase chain reaction (PCR). Illumina HiSeq 4000 was used for Pair-End 101bp sequencing.