title: GSM4494535: CITCO_replicate 2; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA (RIN > 9 by Bioanalyzer; Agilent Technologies) was isolated using RNeasy columns with on-column DNase treatment (QIAGEN) and processed with the NEBNext Ultra II ribosomal (r)RNA-depletion module Human/Mouse/Rat (New England BioLabs Inc.). Using 400 ng total RNA as input rRNA-depleted RNA was processed using the NEBNext Ultra II directional RNA kit for Illumina (New England BioLabs Inc.) to generate strand-specific libraries. Final libraries were amplified by PCR (10-cycles), size selected using beads to enrich for fragments > 400 bp and purified using Ampure XP beads (Beckman Coulter). Final libraries were validated on the Agilent 2100 Bioanalyzer DNA chips and quantified on the Qubit (Thermo Fisher Scientific). Libraries were pooled at equimolar ratios and loaded onto the Illumina NextSeq500 flow cell and sequenced using 40 bp paired-end chemistry.
identifiers.namespace: GEO
identifiers.id: GSM4494535