Paired-end (PE) RNA-seq library
A paired-end (PE) RNA-seq library was prepared using a pooled RNA sample and a non-stranded, massively-parallel cDNA sequencing (mRNA-Seq) protocol with the TruSeq mRNA prep Kit (Cat. No. RS-122-2101) from Illumina, Inc. (San Diego, CA, USA). The ligation step in the protocol was optimized by diluting the adapters concentration to increase ligation efficiency (> 94 %), and the number of PCR cycles was reduce to avoid PCR duplication artefacts as well as primer dimers in the final library. The fluorometric based QuantiFluordsDNA System (Promega GmbH, Mannheim, Germany) was used for accurate quantitation of the cDNA library. The size of the final cDNA library was determined by using the dsDNA 905 Reagent Kit (Agilent Technologies, Inc., Santa Clara, CA, USA) with sizing range of 35-500 bp and resolution of 3-5 bp at 300 bp on average. The PE library was sequenced in two flowcell lanes on the Illumina HiSeq2500 with a rapid mode and 2  250 cycles.
SUB6541048
PE_RNA