GSM5741220: Ctrl_3; Homo sapiens; RNA-Seq
Total RNA was isolated using Direct-zol RNA extraction kit from Zymo Research Library preparation was performed from 200ng total RNA using the TruSeq Stranded mRNA Library Kit (Illumina, 20020594) and the TruSeq RNA UD Indexes (Illumina, 20020590). 15 cycles of PCR were performed. Libraries were quality-checked on a Fragment Analyzer (Advanced Analytical) using the Standard Sensitivity NGS Fragment Analysis Kit (Advanced Analytical, DNF-474) revealing excellent quality of libraries (average concentration: 145±43 nmol/L, average library size: 339±8 base pairs). Samples were pooled to equal molarity. The pool was quantified by Fluorometry using the QuantiFluor ONE dsDNA System (Promega, E4871). Libraries were sequenced Single-reads 76 bases using a NextSeq 500 High Output Kit 75-cycles (Illumina, 20024906) loaded at 1.8pM and including 1% PhiX.