Measuring technical variability in illumina DNA methylation microarrays.

DNA methylation microarrays have become a widely used tool for investigating epigenetic modifications in various aspects of biomedical research. However, technical variability in methylation data poses challenges for downstream applications such as predictive modeling of health and disease. In this...

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Main Authors: Anderson A Butler, Jason J Kras, Karolina P Chwalek, Enrique I Ramos, Isaac J Bishof, David S Vogel, Daniel L Vera
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2025-01-01
Series:PLoS ONE
Online Access:https://doi.org/10.1371/journal.pone.0326337
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author Anderson A Butler
Jason J Kras
Karolina P Chwalek
Enrique I Ramos
Isaac J Bishof
David S Vogel
Daniel L Vera
author_facet Anderson A Butler
Jason J Kras
Karolina P Chwalek
Enrique I Ramos
Isaac J Bishof
David S Vogel
Daniel L Vera
author_sort Anderson A Butler
collection DOAJ
description DNA methylation microarrays have become a widely used tool for investigating epigenetic modifications in various aspects of biomedical research. However, technical variability in methylation data poses challenges for downstream applications such as predictive modeling of health and disease. In this study, we measure the impact of common sources of technical variability in Illumina DNA methylation microarray data, with a specific focus on positional biases inherent within the microarray technology. By utilizing a dataset comprised of multiple, highly similar technical replicates, we identified a chamber number bias, with different chambers of the microarray exhibiting systematic differences in fluorescence intensities (FI) and their derived methylation beta values, which are only partially corrected for by existing preprocessing methods and demonstrate that this positional bias can lead to false positive results during differential methylation testing. Additionally, our investigation identified outliers in low-level fluorescence data which might play a role in contributing to predictive error in computational models of health-relevant traits such as age.
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institution Kabale University
issn 1932-6203
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publishDate 2025-01-01
publisher Public Library of Science (PLoS)
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series PLoS ONE
spelling doaj-art-8e3126861bff4857bbcdb4dce63df8dd2025-08-20T03:27:52ZengPublic Library of Science (PLoS)PLoS ONE1932-62032025-01-01207e032633710.1371/journal.pone.0326337Measuring technical variability in illumina DNA methylation microarrays.Anderson A ButlerJason J KrasKarolina P ChwalekEnrique I RamosIsaac J BishofDavid S VogelDaniel L VeraDNA methylation microarrays have become a widely used tool for investigating epigenetic modifications in various aspects of biomedical research. However, technical variability in methylation data poses challenges for downstream applications such as predictive modeling of health and disease. In this study, we measure the impact of common sources of technical variability in Illumina DNA methylation microarray data, with a specific focus on positional biases inherent within the microarray technology. By utilizing a dataset comprised of multiple, highly similar technical replicates, we identified a chamber number bias, with different chambers of the microarray exhibiting systematic differences in fluorescence intensities (FI) and their derived methylation beta values, which are only partially corrected for by existing preprocessing methods and demonstrate that this positional bias can lead to false positive results during differential methylation testing. Additionally, our investigation identified outliers in low-level fluorescence data which might play a role in contributing to predictive error in computational models of health-relevant traits such as age.https://doi.org/10.1371/journal.pone.0326337
spellingShingle Anderson A Butler
Jason J Kras
Karolina P Chwalek
Enrique I Ramos
Isaac J Bishof
David S Vogel
Daniel L Vera
Measuring technical variability in illumina DNA methylation microarrays.
PLoS ONE
title Measuring technical variability in illumina DNA methylation microarrays.
title_full Measuring technical variability in illumina DNA methylation microarrays.
title_fullStr Measuring technical variability in illumina DNA methylation microarrays.
title_full_unstemmed Measuring technical variability in illumina DNA methylation microarrays.
title_short Measuring technical variability in illumina DNA methylation microarrays.
title_sort measuring technical variability in illumina dna methylation microarrays
url https://doi.org/10.1371/journal.pone.0326337
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