GSM4082148: Sample 3 CEM.cHSF1 water 18h; Homo sapiens; RNA-Seq
Cellular RNA was extracted using the Qiagen RNeasy Plus Mini Kit and QIAshredder homogenization columns. For RNAseq, RNA integrity and concentration were checked on a Fragment Analyzer (Advanced Analytical) and RNA-seq libraries were prepared using the Kapa mRNA HyperPrep RNA-seq library construction kit (Kapa/Roche), with 7 min fragmentation at 94 °C and 15 PCR cycles of final amplification and duplex barcoding. Libraries were quantified using the Fragment Analyzer (Advanced Analytical) and qPCR before being loaded for single-end sequencing using the Illumina HiSeq 2000 for 40 nucleotides in High Output mode. polyA-based mRNA capture using RNA-Kapa with TruSeq strand-specific strategy.