title: A simple modification to the luminometric methylation assay to control for the effects of DNA fragmentation.
abstract: 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.
authors.affiliation: Department of Psychology, Bogazici University, Istanbul, Turkey.|Center for Life Sciences and Technologies, Bogazici University, Istanbul, Turkey.
authors.forename: Elif Aysimi
authors.initials: EA
authors.lastname: Duman
authors.affiliation: Ludwig Institute for Cancer Research Ltd, University of Oxford, Oxford, U.K.
authors.forename: Skirmantas
authors.initials: S
authors.lastname: Kriaucionis
authors.affiliation: Biology Department, Brookhaven National Laboratory, Upton, NY.
authors.forename: John J
authors.initials: JJ
authors.lastname: Dunn
authors.affiliation: Department of Pathology, Stony Brook University, Stony Brook, NY.
authors.forename: Eli
authors.initials: E
authors.lastname: Hatchwell
mesh_terms: D001681:Biological Assay; D004247:DNA / Q000032:analysis* / Q000235:genetics; D053938:DNA Fragmentation; D019175:DNA Methylation; D004262:DNA Restriction Enzymes / Q000378:metabolism; D006801:Humans
chemical_list: D004247:DNA; D004262:DNA Restriction Enzymes
keywords: DNA integrity; epigenetics; global DNA methylation; luminometric methylation assay