title: GSM5652459: P10_Primary_area_2; Homo sapiens; RNA-Seq
library_construction_protocol: Simultaneous DNA and total RNA extraction using the AllPrep DNA/RNA FFPE Kit (Qiagen, Hilden, Germany) according to the manufacturer's protocol. Prior to sequencing, RNA integrity was assessed using the Agilent Bioanalyzer RNA 6000 Nano Kit (Agilent Technologies, Waldbronn, Germany) by calculating the percentage of fragments > 200 nucleotides (DV200). Sequencing libraries were prepared with 100 ng total RNA using the QuantSeq 3'-RNA-Seq Library Prep Kit FWD for Illumina with i5 6 nt Dual Indexing(Lexogen GmbH, Vienna, Austria) according to the manufacturer's instructions for dual-indexing and low quantity samples. For library amplification, PCR cycles were determined using the PCR Add-on Kit for Illumina (Lexogen). Quantity and quality of sequencing libraries were assessed using the Quanti-iT PicoGreen dsDNA Assay Kit (ThermoScientific) and the Bioanalyzer High Sensitivity DNA Analysis Kit (Agilent Technologies). Libraries were sequenced in 150bp paired-end mode on a HiSeq4000 sequencer (Illumina, Berlin, Germany). The pool of individually barcoded libraries was distributed across the lanes of the same flow-cell aiming for approximately ten million paired-end reads per sample. 150bp paired-end