title: GSM4066075: ext_L13372.1_301; Homo sapiens; RNA-Seq
library_construction_protocol: Cellular RNA was isolated using the NucleoSpin® RNA Isolation kit (Macherey-Nagel, Düren, Germany) according to manufacturer's instructions. For each experimental condition, three biological replicates were analyzed. After initial quality control using Agilent's Bioanalyzer, sequencing libraries were prepared from 500ng of total RNA per sample following Roche's stranded “KAPA RNA HyperPrep” library preparation protocol for single indexed Illumina libraries: First the polyA-RNA fraction was enriched using oligo-dT-probed paramagnetic beads. Enriched RNA was heat-fragmented and subjected to first strand synthesis using random priming. The second strand was synthesized incorporating dUTP instead of dTTP to preserve strand information. After A-tailing Illumina sequencing compatible adapters were ligated. Following bead-based clean-up steps the libraries were amplified using 10 cycles of PCR. Library quality and size was checked with qBit, Agilent Bioanalyzer and qPCR. Sequencing was carried out on an Illumina HiSeq 4000 system in PE75bp mode yielding between 15-23 million fragments per sample.
attributes.tag: GEO Accession
attributes.value: GSM4066075