SIRV Validation Experiment NGS1.61, RC0 sample, TopHat2 pipeline
The experiments were carried out in technical triplicates. From 1 microgram sample RC-0 NGS libraries  were prepared by Fasteris SA (Plan-les-Ouates, CH), using the TruSeq stranded mRNA library preparation kit with polyA selection (Illumina). All primary library templates were amplified with 14 PCR cycles to reach reproducible concentrations, 155+/-45 nM, and insert sizes, 165+/-10 bp.  One equimolar lane-mix was sequenced at the CSF Campus Science Support Facilities GmbH (CSF, Vienna, AUT) in PE125 mode on in one lane of a HiSeq5A flowcell using V4 chemistry. The run yielded 305 M raw reads, 293 M after demultiplexing and trimming which corresponds to a read depth of 32.6+/-2.2 M reads per library.