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BMC Genomics

, 9:59

First Online: 31 January 2008Received: 08 May 2007Accepted: 31 January 2008

Abstract

BackgroundDNA methylation based techniques are important tools in both clinical diagnostics and therapeutics. But most of these methods only analyze a few CpG sites in a target region. Indeed, difference of site-specific methylation may also lead to a change of methylation density in many cases, and it has been found that the density of methylation is more important than methylation of single CpG site for gene silencing.

ResultsWe have developed a novel approach for quantitative analysis of CpG methylation density on the basis of microarray-based hybridization and incorporation of Cy5-dCTP into the Cy3 labeled target DNA by using Taq DNA Polymerase on microarray. The quantification is achieved by measuring Cy5-Cy3 signal ratio which is proportional to methylation density. This methylation-sensitive technique, termed RMEAM regional methylation elongation assay on microarray, provides several advantages over existing methods used for methylation analysis. It can determine an exact methylation density of the given region, and has potential of high throughput. We demonstrate a use of this method in determining the methylation density of the promoter region of the tumor-related gene MLH1, TERT and MGMT in colorectal carcinoma patients.

ConclusionThis technique allows for quantitative analysis of regional methylation density, which is the representative of all allelic methylation patterns in the sample. The results show that this technique has the characteristics of simplicity, rapidness, specificity and high-throughput.

Electronic supplementary materialThe online version of this article doi:10.1186-1471-2164-9-59 contains supplementary material, which is available to authorized users.

Dingdong Zhang, Yan Wang contributed equally to this work.

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Autor: Dingdong Zhang - Yan Wang - Yunfei Bai - Qinyu Ge - Yingjuan Qiao - Junfeng Luo - Chao Jia - Zuhong Lu

Fuente: https://link.springer.com/article/10.1186/1471-2164-9-59







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