dupRadar: a Bioconductor package for the assessment of PCR artifacts in RNA-Seq data
Abstract
BackgroundPCR clonal artefacts originating from NGS library preparation can affect both genomic as well as RNA-Seq applications when protocols are pushed to their limits. In RNA-Seq however the artifactual reads are not easy to tell apart from normal read duplication due to natural over-sequencing of highly expressed genes. Especially when working with little input material or single cells assessing the fraction of duplicate reads is an important quality control step for NGS data sets. Up to now there are only tools to calculate the global duplication rates that do not take into account the effect of gene expression levels which leaves them of limited use for RNA-Seq data.\n\nResultsHere we present the tool dupRadar, which provides an easy means to distinguish artefactual from natural duplicate reads in RNA-Seq data. dupRadar assesses the fraction of duplicate reads per gene dependent on the expression level. Apart from the Bioconductor package dupRadar we provide shell scripts for easy integration into processing pipelines.\n\nConclusionsThe Bioconductor package dupRadar offers straight-forward methods to assess RNA-Seq datasets for quality issues with PCR duplicates. It is aimed towards simple integration into standard analysis pipelines as a default QC metric that is especially useful for low-input and single cell RNA-Seq data sets.
02 /Specification & Classification
Click any tag to browse matching papersReproducibility ledger
Nobody has reported trying to reproduce this paper yet. Did you replicate an experiment or run the code? Your report saves the next lab a week.
Sign in to file a reproducibility report.
Related papers
- Effects of duplicated mapped read PCR artifacts on RNA-seq differential expression analysis based on qRNA-seq
Anna C. Salzberg, Jiafen Hu, … Yuka Imamura Kawasawa·bioRxiv·Apr 13, 2018
- The impact of PCR duplication on RNA-seq data generated using NovaSeq 6000, NovaSeq X, AVITI and G4 sequencers.
Natalia Zajac, Ioannis S. Vlachos, … Catharine Aquino·Genome BiologyPeer-reviewed·May 28, 2025
- Elimination of PCR duplicates in RNA-seq and small RNA-seq using unique molecular identifiers
Yu Fu, Pei‐Hsuan Wu, … Zhiping Weng·BMC GenomicsPeer-reviewed·Jul 12, 2018
- The impact of amplification on differential expression analyses by RNA-seq
Swati Parekh, Christoph Ziegenhain, … Ines Hellmann·Scientific ReportsPeer-reviewed·May 9, 2016
- SARTools: a DESeq2- and edgeR-based R pipeline for comprehensive differential analysis of RNA-Seq data
Hugo Varet, Jean-Yves Coppée, Marie-Agnès Dillies·bioRxiv·Sep 26, 2015
- On the causes, consequences, and avoidance of PCR duplicates: towards a theory of library complexity
Nicolas C. Rochette, Angel G. Rivera‐Colón, … Julian Catchen·Molecular Ecology ResourcesPeer-reviewed·Apr 16, 2023