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SIRVs: Spike-In RNA Variants as External Isoform Controls in RNA-Sequencing

Lukas Paul, Petra Kubala, Gudrun Horner, Michael Ante, Igor Holländer, Seitz Alexander, Torsten Reda
doi: https://doi.org/10.1101/080747
Lukas Paul
Lexogen GmbH, Campus Vienna Biocenter 5, 1030 Vienna, Austria
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Petra Kubala
Lexogen GmbH, Campus Vienna Biocenter 5, 1030 Vienna, Austria
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Gudrun Horner
Lexogen GmbH, Campus Vienna Biocenter 5, 1030 Vienna, Austria
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Michael Ante
Lexogen GmbH, Campus Vienna Biocenter 5, 1030 Vienna, Austria
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Igor Holländer
Lexogen GmbH, Campus Vienna Biocenter 5, 1030 Vienna, Austria
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Seitz Alexander
Lexogen GmbH, Campus Vienna Biocenter 5, 1030 Vienna, Austria
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Torsten Reda
Lexogen GmbH, Campus Vienna Biocenter 5, 1030 Vienna, Austria
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Abstract

Spike-In RNA variants (SIRVs) enable for the first time the validation of RNA sequencing workflows using external isoform transcript controls. 69 transcripts, derived from seven human model genes, cover the eukaryotic transcriptome complexity of start- and end-site variations, alternative splicing, overlapping genes, and antisense transcription in a condensed format. Reference RNA samples were spiked with SIRV mixes, sequenced, and exemplarily four data evaluation pipelines were challenged to account for biases introduced by the RNA-Seq workflow. The deviations of the respective isoform quantifications from the known inputs allow to determine the comparability of sequencing experiments and to extrapolate to which degree alterations in an RNA-Seq workflow affect gene expression measurements. The SIRVs as external isoform controls are an important gauge for inter-experimental comparability and a modular spike-in contribution to clear the way for diagnostic RNA-Seq applications.

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Posted October 13, 2016.
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SIRVs: Spike-In RNA Variants as External Isoform Controls in RNA-Sequencing
Lukas Paul, Petra Kubala, Gudrun Horner, Michael Ante, Igor Holländer, Seitz Alexander, Torsten Reda
bioRxiv 080747; doi: https://doi.org/10.1101/080747
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SIRVs: Spike-In RNA Variants as External Isoform Controls in RNA-Sequencing
Lukas Paul, Petra Kubala, Gudrun Horner, Michael Ante, Igor Holländer, Seitz Alexander, Torsten Reda
bioRxiv 080747; doi: https://doi.org/10.1101/080747

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