Accurate quantification of DNA methylation using combined bisulfite restriction analysis coupled with the Agilent 2100 Bioanalyzer platform.
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.
Brena
Romulo Martin
RM
Department of Molecular Genetics, The Ohio State University, Columbus, OH, USA.
Auer
Herbert
H
Kornacker
Karl
K
Hackanson
Björn
B
Raval
Aparna
A
Byrd
John C
JC
Plass
Christoph
C
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
D013447:Sulfites; D000077209:Decitabine; D004262:DNA Restriction Enzymes; D001374:Azacitidine