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Tackling the widespread and critical impact of batch effects in high-throughput data
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High-throughput technologies are widely used, for example to assay genetic variants, gene and protein expression, and epigenetic modifications. One often overlooked complication with such studies is batch effects, which occur because measurements are affected by laboratory conditions, reagent lots and personnel differences. This becomes a major problem when batch effects are correlated with an outcome of interest and lead to incorrect conclusions. Using both published studies and our own analyses, we argue that batch effects (as well as other technical and biological artefacts) are widespread and critical to address. We review experimental and computational approaches for doing so.
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“These batch effects make it difficult to draw conclusions from data across experimental batches (Leek et al., 2010; Parker and Leek, 2012; Soneson et al., 2014; Nygaard et al., 2016; Caicedo et al., 2017).”
From WILDS: A Benchmark of in-the-Wild Distribution Shifts (2020) · cites this paper as Leek et al., 2010
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“Several methods have been suggested to adjust for batch effects, including recommendations for experimental designs and statistical approaches (see for example Leek et al., 2010).”
From The applied implications of epigenetics in anxiety, affective and stress-related disorders - A review and synthesis on psychosocial stress, psychotherapy and prevention (2020) · cites this paper as Leek et al., 2010
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“Despite attempts to carefully control experimental variables such as temperature, humidity, and reagent concentration, measurements from these screens are confounded by technical artifacts that arise from differences in the execution of each batch.”
From WILDS: A Benchmark of in-the-Wild Distribution Shifts (2020) · cites this paper as Leek et al., 2010
In the citing paper
“Batch effects and technical variation due to unknown (unmodeled) confounders are common in both scRNA-seq and bulk RNA-seq.”
From Revealing the vectors of cellular identity with single-cell genomics (2016) · cites this paper as [45]
In the citing paper
“Notably, edges are susceptible to ascertainment biases52,214,215 and confounding factors that can induce spurious relationships178 (FIG. 2b).”
From Systems biology and gene networks in neurodevelopmental and neurodegenerative disorders (2015) · cites this paper as [178]
In the citing paper
“Optimal methodological choices and study designs ensure that the biological signals from the main factors are not confounded by variation from unwanted factors178.”
From Systems biology and gene networks in neurodevelopmental and neurodegenerative disorders (2015) · cites this paper as [178]
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“there are no batch normalization approaches for microbiome data, unlike in transcriptional or genetic meta-analyses26.”
From Assessment of variation in microbial community amplicon sequencing by the Microbiome Quality Control (MBQC) project consortium (2017) · cites this paper as [26]
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Metadata
- Authors
- Leek, J. T., Scharpf, R. B., Bravo, H. C., Simcha, D., Langmead, B., Johnson, W. E., Geman, D., Baggerly, K., & Irizarry, R. A.
- Affiliations
- Johns Hopkins University (US)
- Brigham Young University (US)
- The University of Texas MD Anderson Cancer Center (US)
- Corresponding author
- Irizarry, Rafael A.
- Venue
- Nature Reviews Genetics
- Published
- 2010-09-14
- Volume
- 11
- Issue
- 10
- Pages
- 733-739
- ISSN
- 1471-0056 (ISSN-L)
- 1471-0064
- PMC
- PMC3880143
- OpenAlex
- W2096192437
- Cited by
- 2,196
- References
- 30
- Citations by year
- 2026: 235 · 2025: 166 · 2024: 159 · 2023: 125 · 2022: 178 · 2021: 178 · 2020: 167 · 2019: 138 · 2018: 152 · 2017: 130 · 2016: 133 · 2015: 107 · 2014: 100 · 2013: 95 · 2012: 70
- FWCI
- 6.29
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature Reviews Genetics (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- Juvenile Diabetes Research Foundation International
- Wellcome Trust
- National Institute for Health and Care Research
- National Institutes of Health (5u41hg004059-15)
- Cambridge Institute for Medical Research, University of Cambridge (079895)
- NIHR Cambridge Biomedical Research Centre
- National Human Genome Research Institute (HG005220, U41-HG004059, HG004059, R00HG005015, R01HG005220, P41HG004059, U24HG004059)
- National Institute of General Medical Sciences (GM0083084, R01GM083084)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Gene expression and cancer classification
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Bioinformatics and Genomic Networks
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Single-cell and spatial transcriptomics
- Keywords
- batch effects, high-throughput omics data, confounding, gene expression, protein expression, epigenetic modifications, laboratory conditions
- MeSH terms
- Biotechnology, Periodicals as Topic, Research Design, Sequence Analysis, DNA, Computational Biology, Oligonucleotide Array Sequence Analysis, Genomics
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