Exemplary multiplex bisulfite amplicon data used to demonstrate the utility of Methpat

Wong, Nicholas C., Pope, Bernard J., Candiloro, Ida, Korbie, Darren, Trau, Matt, Wong, Stephen Q., Mikeska, Thomas, van Denderen, Bryce J.W., Thompson, Erik W., Eggers, Stefan, Doyle, Stephen R., & Dobrovic, Alexander (2015) Exemplary multiplex bisulfite amplicon data used to demonstrate the utility of Methpat. Gigascience, 4, Article Number-55.

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Abstract

BACKGROUND: DNA methylation is a complex epigenetic marker that can be analyzed using a wide variety of methods. Interpretation and visualization of DNA methylation data can mask complexity in terms of methylation status at each CpG site, cellular heterogeneity of samples and allelic DNA methylation patterns within a given DNA strand. Bisulfite sequencing is considered the gold standard, but visualization of massively parallel sequencing results remains a significant challenge. FINDINGS: We created a program called Methpat that facilitates visualization and interpretation of bisulfite sequencing data generated by massively parallel sequencing. To demonstrate this, we performed multiplex PCR that targeted 48 regions of interest across 86 human samples. The regions selected included known gene promoters associated with cancer, repetitive elements, known imprinted regions and mitochondrial genomic sequences. We interrogated a range of samples including human cell lines, primary tumours and primary tissue samples. Methpat generates two forms of output: a tab-delimited text file for each sample that summarizes DNA methylation patterns and their read counts for each amplicon, and a HTML file that summarizes this data visually. Methpat can be used with publicly available whole genome bisulfite sequencing and reduced representation bisulfite sequencing datasets with sufficient read depths. CONCLUSIONS: Using Methpat, complex DNA methylation data derived from massively parallel sequencing can be summarized and visualized for biological interpretation. By accounting for allelic DNA methylation states and their abundance in a sample, Methpat can unmask the complexity of DNA methylation and yield further biological insight in existing datasets.

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2 citations in Scopus
1 citations in Web of Science®
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ID Code: 97717
Item Type: Journal Article
Refereed: Yes
Keywords: Cell Line, *DNA Methylation, *High-Throughput Nucleotide Sequencing, Humans, Neoplasms/genetics, Organ Specificity, Sequence Analysis, DNA/*methods, *Software, Bisulfite sequencing, Cancer, DNA methylation, Epialleles, Epigenetics, Pcr, Visualization
DOI: 10.1186/s13742-015-0098-x
ISSN: 2047-217x
Subjects: Australian and New Zealand Standard Research Classification > MEDICAL AND HEALTH SCIENCES (110000)
Divisions: Current > Schools > School of Biomedical Sciences
Current > QUT Faculties and Divisions > Faculty of Health
Current > Institutes > Institute of Health and Biomedical Innovation
Copyright Owner: 2015 Wong et al. Open Access
Copyright Statement: This article is distributed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and
reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to
the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver
(http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
Deposited On: 31 Jul 2016 23:15
Last Modified: 02 Aug 2016 04:13

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