A novel real-time PCR assay for quantitative analysis of methylated alleles (QAMA): analysis of the retinoblastoma locus
作者： Michael Zeschnigk , Stefan Böhringer , Elizabeth Ann Price , Zerrin Onadim , Lars Maßhöfer
DOI: 10.1093/NAR/GNH122
关键词: DNA methylation 、 Illumina Methylation Assay 、 Biology 、 CpG site 、 Genetics 、 Polymerase chain reaction 、 Methylation 、 TaqMan 、 Single-nucleotide polymorphism 、 Methylated DNA immunoprecipitation
摘要: Altered methylation patterns have been found to play a role in developmental disorders, cancer and aging. Increasingly, changes DNA are used as molecular markers of disease. Therefore, there is need for reliable easy use techniques detect measure research routine diagnostics. We established novel quantitative analysis methylated alleles (QAMA) which essentially major improvement over previous method based on real-time PCR (MethyLight). This bisulfite-treated DNA. A significant advantage conventional MethyLight gained by the TaqMan probes minor groove binder (MGB) technology. Their improved sequence specificity facilitates relative quantification unmethylated that simultaneously amplified single tube. allows precise measurement ratio versus cuts down potential sources inter-assay variation. fewer control assays required. this technical approach identify hypermethylation CpG island located promoter region retinoblastoma (RB1) gene QAMA fast quantity improves handling diagnostic analysis. Moreover, simplified reaction setup robustness inherent tube assay high-throughput Because high MGB technology widely discriminate nucleotide polymorphisms, potentially can be status dinucleotides.
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