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Methylation-sensitive high resolution melting (MS-HRM): a new approach for sensitive and high-throughput assessment of methylation
Open accessAbstract
In this article, we show that high resolution melting analysis (HRM) is a sensitive and specific method for the detection of methylation. Methylated DNA and unmethylated DNA acquire different sequences after bisulphite treatment resulting in PCR products with markedly different melting profiles. We used PCR to amplify both methylated and unmethylated sequences and assessed HRM for the determination of the methylation status of the MGMT promoter region. Reconstruction experiments showed that MGMT methylation could be detected at levels as low as 0.1%. Moreover, MS-HRM allows for estimation of the methylation level by comparing the melting profiles of unknown PCR products to the melting profiles of PCR products derived from standards with a known unmethylated to methylated template ratio. We used MS-HRM for the analysis of eight cell lines of known methylation status and a panel of colorectal cancer specimens. The simplicity and high reproducibility of the MS-HRM protocol makes MS-HRM the method of choice for methylation assessment in many diagnostic and research applications.
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Cited by
High-Resolution DNA Melting Analysis for Simple and Efficient Molecular Diagnostics
Reed, G. H., Kent, J. O., & Wittwer, C. T. (2007) · Pharmacogenomics · doi:10.2217/14622416.8.6.597 · Cited by 678
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Bidwell, B. N., Slaney, C. Y., Withana, N. P., Forster, S., Cao, Y., Loi, S., Andrews, D., Mikeska, T., Mangan, N. E., Samarajiwa, S. A., et al. (2012) · Nature Medicine · doi:10.1038/nm.2830 · Cited by 487
Osterlehner, A., Simmeth, S., & Göpfert, U. (2011) · Biotechnology and Bioengineering · doi:10.1002/bit.23216 · Cited by 131
Kobayashi, N., Shinagawa, S., Niimura, H., Kida, H., Nagata, T., Tagai, K., Shimada, K., Oka, N., Shikimoto, R., Noda, Y., et al. (2020) · Scientific Reports · doi:10.1038/s41598-020-69248-9 · Cited by 23
Metadata
- Authors
- Wojdacz, T. K., & Dobrovic, A.
- Venue
- Nucleic Acids Research
- Publisher
- Oxford University Press (OUP)
- Published
- 2007-03-01
- Volume
- 35
- Issue
- 6
- Pages
- e41-e41
- ISSN
- 0305-1048 (ISSN-L)
- 1362-4962
- OpenAlex
- W2098442878
- Cited by
- 551
- References
- 0
- FWCI
- 8.62
- Type
- journal article
- Language
- en
- Open access
- gold
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- Wojdacz, T. K., & Dobrovic, A. (2007). Methylation-sensitive high resolution melting (MS-HRM): a new approach for sensitive and high-throughput assessment of methylation. Nucleic Acids Research. https://doi.org/10.1093/nar/gkm013
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