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Melting Curve Assays for DNA Methylation Analysis

  • Tomasz K. Wojdacz
  • Alexander Dobrovic
Part of the Methods in Molecular Biology book series (MIMB, volume 507)

Abstract

The ability of sodium bisulfite to modify cytosines in a methylation-dependent manner allows the conservation of DNA methylation information during PCR amplification. PCR products amplified from bisulfite-modified DNA have significantly different base compositions according to whether they originate from methylated or unmethylated variants of the target template. Different base compositions give rise to different thermal properties of the PCR products. Hence, melting analysis of amplification products in methylation studies allows the determination of whether the PCR products originate from methylated or unmethylated templates. Here, we briefly review recent advances in methodologies based on melting analyses of PCR products derived from bisulfite-modified templates and provide a methodology for methylation-sensitive high-resolution melting.

Keywords

Methylation melting curve sodium bisulfite high-resolution melting PCR bias, Methylation-sensitive high-resolution melting (MS-HRM) 

Notes

Acknowledgements

Tomasz K. Wojdacz would like to thank Dr Lise Lotte Hansen for the greatly appreciated support and always good advice. The Danish Medical Research Council and Harboefonden supported financially work of T K Wojdacz. Alexander Dobrovic was supported by grants from the National Health and Medical Research Council of Australia, the Cancer Council of Victoria and the US Department of Defense Breast Cancer Research Program. We also would like to thank Dr. Michael Hoffmann and Roche Diagnostics, Penzberg, Germany for help with the publication of this chapter.

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Tomasz K. Wojdacz
    • 1
  • Alexander Dobrovic
    • 2
    • 3
  1. 1.Institute of Human GeneticsUniversity of AarhusDenmark
  2. 2.Molecular Pathology Research and Development Laboratory, Department of PathologyPeter MacCallum Cancer CentreAustralia
  3. 3.Department of PathologyUniversity of MelbourneAustralia

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