Accurate quantification of DNA methylation using combined bisulfite restriction analysis coupled with the Agilent 2100 Bioanalyzer platform | Zendy

Romulo Martin Brena | Zendy; Herbert Auer | Zendy; Karl Kornacker | Zendy; Björn Hackanson | Zendy; Aparna Raval | Zendy; John C. Byrd | Zendy; Christoph Plass | Zendy

Open Access

Accurate quantification of DNA methylation using combined bisulfite restriction analysis coupled with the Agilent 2100 Bioanalyzer platform

Author(s) -

Romulo Martin Brena,

Herbert Auer,

Karl Kornacker,

Björn Hackanson,

Aparna Raval,

John C. Byrd,

Christoph Plass

Publication year - 2006

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/gnj017

Subject(s) - dna methylation , bisulfite , biology , methylation , illumina methylation assay , bisulfite sequencing , epigenetics , dna , microbiology and biotechnology , methylated dna immunoprecipitation , cytosine , computational biology , gene , genetics , gene expression

DNA methylation is the best-studied epigenetic modi- fication and describes the conversion of cytosine to 5-methylcytosine. The importance of this phe- nomenon 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 challen- ging. In this report, we describe Bio-COBRA (com- bined 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.

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