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Variations in DNA methylation have been implicated in a number of disorders. Changes in global DNA methylation levels have long been associated with various types of cancer. One of the recently described methods for determining global DNA methylation levels is the LUminometric Methylation Assay (LUMA), which utilizes methylation sensitive and insensitive restriction endonucleases and pyrosequencing technology for quantification. Here we provide evidence suggesting that the global methylation level reported by LUMA is affected by the integrity of the DNA being analyzed. The less intact the DNA, the lower the global methylation levels reported by LUMA. In order to overcome this problem, we propose the use of undigested DNA alongside digested samples. Finally, we demonstrate that this results in a more accurate assessment of global DNA methylation levels.

Original publication

DOI

10.2144/000114290

Type

Journal article

Journal

Biotechniques

Publication Date

05/2015

Volume

58

Pages

262 - 264

Keywords

DNA integrity, epigenetics, global DNA methylation, luminometric methylation assay, Biological Assay, DNA, DNA Fragmentation, DNA Methylation, DNA Restriction Enzymes, Humans