Cited by 113Molecular Biology Techniques and ApplicationsMeta-analysis and systematic reviewsClinical Laboratory Practices and Quality Control

Talking the talk, but not walking the walk: RT-qPCR as a paradigm for the lack of reproducibility in molecular research

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Bustin, Stephen, Nolan, Tania·2017·European Journal of Clinical Investigation

Abstract

Poorly executed and inadequately reported molecular measurement methods are amongst the causes underlying the lack of reproducibility of much biomedical research. Although several high impact factor journals have acknowledged their past failure to scrutinise adequately the technical soundness of manuscripts, there is a perplexing reluctance to implement basic corrective measures. The reverse transcription real-time quantitative PCR (RT-qPCR) is probably the most straightforward measurement technique available for RNA quantification and is widely used in research, diagnostic, forensic and biotechnology applications. Despite the impact of the minimum information for the publication of quantitative PCR experiments (MIQE) guidelines, which aim to improve the robustness and the transparency of reporting of RT-qPCR data, we demonstrate that elementary protocol errors, inappropriate data analysis and inadequate reporting continue to be rife and conclude that the majority of published RT-qPCR data are likely to represent technical noise.

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Crossref · 10.1111/eci.12801

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Authors
Bustin, S., & Nolan, T.
Affiliations
  • Anglia Ruskin University (GB)
  • University of Manchester (GB)
Corresponding author
Bustin, Stephen
Venue
European Journal of Clinical Investigation
Publisher
Wiley
Published
2017-09-02
Volume
47
Issue
10
Pages
756-774
ISSN
0014-2972 (ISSN-L)
OpenAlex
W2743105407
Cited by
113
References
165
Citations by year
2026: 7 · 2025: 15 · 2024: 14 · 2023: 8 · 2022: 16 · 2021: 13 · 2020: 14 · 2019: 17 · 2018: 8 · 2017: 1
FWCI
1.02
Type
journal article
Language
en
Open access
Locations
  • European Journal of Clinical Investigation (publishedVersion)
  • PubMed (publishedVersion)
  • Anglia Ruskin Research Online (Anglia Ruskin University) (submittedVersion)
  • Figshare (submittedVersion, cc-by)
  • Research Explorer (The University of Manchester) (submittedVersion)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Molecular Biology Techniques and Applications
  • Social Sciences › Decision Sciences › Statistics, Probability and Uncertainty › Meta-analysis and systematic reviews
  • Health Sciences › Medicine › Physiology › Clinical Laboratory Practices and Quality Control
Keywords
RT-qPCR, reproducibility, MIQE guidelines, reporting quality, technical noise, RNA quantification, data analysis, research integrity
MeSH terms
Humans, RNA, Sensitivity and Specificity, Reproducibility of Results, Biomedical Research, Journal Impact Factor, Real-Time Polymerase Chain Reaction

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