GSM2753390: WT SETMAR 4; Homo sapiens; RNA-Seq
Cells were harvested 48 hours post-transfection and RNA was isolated using the QIAGEN RNeasy Mini Kit. DNaseI digestion was performed immediately following RNA isolation using PureLink DNase according to manufacturer instructions, and samples were purified using the QIAGEN RNeasy Mini Kit. 500 nanograms of RNA per sample were then used to prepare a single-indexed strand-specific cDNA library using the TruSeq Stranded mRNA Library Prep Kit (Illumina). The resulting libraries were assessed for quantity and size distribution using Qubit and Agilent 2100 Bioanalyzer. 200 pM pooled libraries were utilized per flowcell for clustering amplification on cBot using HiSeq 3000/4000 PE Cluster Kit and sequenced with 2×75bp paired-end configuration on HiSeq4000 (Illumina) using HiSeq 3000/4000 PE SBS Kit. A Phred quality score (Q score) was used to measure the quality of sequencing. More than 90% of the sequencing reads reached Q30 (99.9% base call accuracy). Sequencing data were first assessed using FastQC for quality control.