Critical evaluation of the Illumina MethylationEPIC BeadChip microarray for whole-genome DNA methylation profiling
Open accessAbstract
BACKGROUND: In recent years the Illumina HumanMethylation450 (HM450) BeadChip has provided a user-friendly platform to profile DNA methylation in human samples. However, HM450 lacked coverage of distal regulatory elements. Illumina have now released the MethylationEPIC (EPIC) BeadChip, with new content specifically designed to target these regions. We have used HM450 and whole-genome bisulphite sequencing (WGBS) to perform a critical evaluation of the new EPIC array platform. RESULTS: EPIC covers over 850,000 CpG sites, including >90 % of the CpGs from the HM450 and an additional 413,743 CpGs. Even though the additional probes improve the coverage of regulatory elements, including 58 % of FANTOM5 enhancers, only 7 % distal and 27 % proximal ENCODE regulatory elements are represented. Detailed comparisons of regulatory elements from EPIC and WGBS show that a single EPIC probe is not always informative for those distal regulatory elements showing variable methylation across the region. However, overall data from the EPIC array at single loci are highly reproducible across technical and biological replicates and demonstrate high correlation with HM450 and WGBS data. We show that the HM450 and EPIC arrays distinguish differentially methylated probes, but the absolute agreement depends on the threshold set for each platform. Finally, we provide an annotated list of probes whose signal could be affected by cross-hybridisation or underlying genetic variation. CONCLUSION: The EPIC array is a significant improvement over the HM450 array, with increased genome coverage of regulatory regions and high reproducibility and reliability, providing a valuable tool for high-throughput human methylome analyses from diverse clinical samples.
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One suggestive CpG (cg04099436) was flagged for cross-reactivity and thus omitted from the results.
“One of the suggestive CpGs (cg04099436) was flagged for cross-reactivity (cf. Lehne et al., 2015; Pidsley et al., 2016) and thus omitted from the results.”
From Epigenome-wide DNA methylation in unipolar depression: predictive biomarker of antidepressant treatment response? (2024) · cites this paper as Pidsley et al., 2016
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- Authors
- Pidsley, R., Zotenko, E., Peters, T. J., Lawrence, M. G., Risbridger, G. P., Molloy, P., Van Djik, S., Muhlhausler, B., Stirzaker, C., & Clark, S. J.
- Affiliations
- Garvan Institute of Medical Research (AU)
- Monash University (AU)
- Commonwealth Scientific and Industrial Research Organisation (AU)
- Corresponding author
- Clark, Susan J.
- ORCID
- Venue
- Genome biology
- Published
- 2016-10-07
- Volume
- 17
- Issue
- 1
- Pages
- 208-208
- ISSN
- 1465-6906 (ISSN-L)
- 1474-760X
- PMC
- PMC5055731
- OpenAlex
- W2529024004
- Cited by
- 1,480
- References
- 42
- Citations by year
- 2026: 122 · 2025: 184 · 2024: 229 · 2023: 201 · 2022: 191 · 2021: 208 · 2020: 134 · 2019: 109 · 2018: 72 · 2017: 27 · 2014: 1 · 2013: 1
- FWCI
- 36.05
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- cc-by
- Locations
- Genome biology (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Monash University Research Portal (Monash University) (acceptedVersion, other-oa)
- UNSWorks (University of New South Wales, Sydney, Australia) (submittedVersion, cc-by-nc-nd)
- Adelaide Research & Scholarship (AR&S) (University of Adelaide) (submittedVersion, cc-by)
- Funders
- National Health and Medical Research Council (1035721, 1102752)
- Cancer Australia (1044458)
- Science and Industry Endowment Fund (AU) (RP03-064)
- Cancer Institute of New South Wales (14/ECF/1-23)
- Prostate Cancer Foundation of Australia (YI0911)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Epigenetics and DNA Methylation
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Gene expression and cancer classification
- Keywords
- DNA methylation profiling, distal regulatory elements, enhancers, genetic variation, reproducibility, differentially methylated probes
- MeSH terms
- Enhancer Elements, Genetic, Humans, Genome, Human, Sequence Analysis, DNA, CpG Islands, DNA Methylation, Oligonucleotide Array Sequence Analysis, High-Throughput Nucleotide Sequencing
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