Helmholtz Gemeinschaft

Search
Browse
Statistics
Feeds

cDNA-detector: detection and removal of cDNA contamination in DNA sequencing libraries

[img]
Preview
PDF (Original Article) - Requires a PDF viewer such as GSview, Xpdf or Adobe Acrobat Reader
2MB
[img] Other (Supplementary Information)
6MB

Item Type:Article
Title:cDNA-detector: detection and removal of cDNA contamination in DNA sequencing libraries
Creators Name:Qi, M. and Nayar, U. and Ludwig, L.S. and Wagle, N. and Rheinbay, E.
Abstract:BACKGROUND: Exogenous cDNA introduced into an experimental system, either intentionally or accidentally, can appear as added read coverage over that gene in next-generation sequencing libraries derived from this system. If not properly recognized and managed, this cross-contamination with exogenous signal can lead to incorrect interpretation of research results. Yet, this problem is not routinely addressed in current sequence processing pipelines. RESULTS: We present cDNA-detector, a computational tool to identify and remove exogenous cDNA contamination in DNA sequencing experiments. We demonstrate that cDNA-detector can identify cDNAs quickly and accurately from alignment files. A source inference step attempts to separate endogenous cDNAs (retrocopied genes) from potential cloned, exogenous cDNAs. cDNA-detector provides a mechanism to decontaminate the alignment from detected cDNAs. Simulation studies show that cDNA-detector is highly sensitive and specific, outperforming existing tools. We apply cDNA-detector to several highly-cited public databases (TCGA, ENCODE, NCBI SRA) and show that contaminant genes appear in sequencing experiments where they lead to incorrect coverage peak calls. CONCLUSIONS: cDNA-detector is a user-friendly and accurate tool to detect and remove cDNA detection in NGS libraries. This two-step design reduces the risk of true variant removal since it allows for manual review of candidates. We find that contamination with intentionally and accidentally introduced cDNAs is an underappreciated problem even in widely-used consortium datasets, where it can lead to spurious results. Our findings highlight the importance of sensitive detection and removal of contaminant cDNA from NGS libraries before downstream analysis.
Keywords:Contamination, Genomics, Software, Quality Control, cDNA
Source:BMC Bioinformatics
ISSN:1471-2105
Publisher:BioMed Central
Volume:22
Number:1
Page Range:611
Date:24 December 2021
Official Publication:https://doi.org/10.1186/s12859-021-04529-2
PubMed:View item in PubMed

Repository Staff Only: item control page

Downloads

Downloads per month over past year

Open Access
MDC Library