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Published online before print October 4, 2007
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Copyright © 2007 American Society for Investigative Pathology
Journal of Molecular Diagnostics, doi:10.2353/jmoldx.2007.070025

Accepted for publication July 3, 2007.

Article

A New Dimethyl Sulfoxide-Based Method for Gene Promoter Methylation Detection

Natalia Kholod@, Jacques Boniver, and Philippe Delvenne

From the Department of Pathology, University of Liege, Liege, Belgium

@ To whom correspondence should be addressed. E-mail: natalia.kholod{at}ulg.ac.be.


  Abstract

The identification of gene promoter methylation is a useful tool for the molecular diagnosis of human diseases. We have developed a new PCR-based technique for detecting the methylation status of CpG islands of gene promoters. This new method, named methyl-sensitive dimethyl sulfoxide-PCR (Ms-DMSO-PCR), is based on the finding that methylated and unmethylated DNAs show a different sensitivity to the amount of DMSO used in the PCR reaction. For the amplification of methylated DNA, more DMSO is required in comparison to unmethylated DNA. This finding resulted in the development of a simple PCR screening of CpG islands with addition of DMSO in the range from 0 to 8% (v/v), and the same pair of primers is sufficient for distinguishing hyper- or hypomethylated gene promoters from normally methylated sequences. This new technique is a one-step procedure and does not require any modifications of DNA or expensive equipment. Therefore, Ms-DMSO-PCR has the potential to be widely used for clinical applications as well in basic research.






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Copyright © 2007 by the American Society for Investigative Pathology and the Association for Molecular Pathology.