title: Accurate quantification of DNA methylation using combined bisulfite restriction analysis coupled with the Agilent 2100 Bioanalyzer platform.
abstract: DNA methylation is the best-studied epigenetic modification and describes the conversion of cytosine to 5-methylcytosine. The importance of this phenomenon is that aberrant promoter hypermethylation is a common occurrence in cancer and is frequently associated with gene silencing. Various techniques are currently available for the analysis of DNA methylation. However, accurate and reproducible quantification of DNA methylation remains challenging. In this report, we describe Bio-COBRA (combined bisulfite restriction analysis coupled with the Agilent 2100 Bioanalyzer platform), as a novel approach to quantitative DNA methylation analysis. The combination of a well-established method, COBRA, which interrogates DNA methylation via the restriction enzyme analysis of PCR-amplified bisulfite treated DNAs, with the Bioanalyzer platform allows for the rapid and quantitative assessment of DNA methylation patterns in large sample sets. The sensitivity and reproducibility of Bio-COBRA make it a valuable tool for the analysis of DNA methylation in clinical samples, which could aid in the development of diagnostic and prognostic parameters with respect to disease detection and management.
authors.affiliation: Department of Molecular Genetics, The Ohio State University, Columbus, OH, USA.
authors.forename: Romulo Martin
authors.initials: RM
authors.lastname: Brena
authors.forename: Herbert
authors.initials: H
authors.lastname: Auer
authors.forename: Karl
authors.initials: K
authors.lastname: Kornacker
authors.forename: Björn
authors.initials: B
authors.lastname: Hackanson
authors.forename: Aparna
authors.initials: A
authors.lastname: Raval
authors.forename: John C
authors.initials: JC
authors.lastname: Byrd
authors.forename: Christoph
authors.initials: C
authors.lastname: Plass
mesh_terms: D001374:Azacitidine / Q000031:analogs & derivatives / Q000494:pharmacology; D045744:Cell Line, Tumor; D018899:CpG Islands; D019175:DNA Methylation; D004262:DNA Restriction Enzymes; D000077209:Decitabine; D046209:Electrophoresis, Microchip / Q000295:instrumentation*; D006801:Humans; D025202:Molecular Diagnostic Techniques; D009369:Neoplasms / Q000175:diagnosis; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D013447:Sulfites
chemical_list: D013447:Sulfites; D000077209:Decitabine; D004262:DNA Restriction Enzymes; D001374:Azacitidine