Open AccessA simple modification to the luminometric methylation assay to control for the effects of DNA fragmentation
Author(s) -
Elif Duman,
Skirmantas Kriaučionis,
John J. Dunn,
Eli Hatchwell
Publication year - 2015
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/000114290
Subject(s) - dna methylation , methylation , microbiology and biotechnology , illumina methylation assay , dna , biology , methylated dna immunoprecipitation , restriction enzyme , pyrosequencing , genetics , gene , gene expression
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.