GSM7143017: DMSO1; Homo sapiens; RNA-Seq
First-strand cDNA synthesis was performed with 200 ng total RNA using anchored oligo-dT and SuperScript III First-Strand Synthesis with SuperMix. Second-strand cDNA synthesis was performed using RNase H, DNA Polymerase I, and Invitrogen Second Strand Buffer. Double-stranded cDNA was purified using DNA Clean & Concentrator-5. Library preparation was performed using the Nextera XT DNA Library Preparation Kit. Libraries were cleaned using Agencourt AMPure XP beads according to manufacturer's instructions. Libraries were evaluated by the High Sensitivity DNA Kit on a 2100 Bioanalyzer. They were then multiplexed and sequenced on an Illumina HiSeq with 50 bp paired-end reads. Reads were aligned to NCBI build 37.2 using Tophat (v2.1.0). Cuffquant and Cuffnorm (Cufflinks v2.2.1) were used to quantify expression levels and calculate normalized FPKM values.