原创力文档a refined, rapid and reproducible high resolution melt (hrm)-based method suitable for quantification of global line-1 repetitive element methylation精制、快速和可再生的高分辨率融化(人力资源管理)的方法适合量化全球第1行重复元素的甲基化.pdfVIP
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a refined, rapid and reproducible high resolution melt (hrm)-based method suitable for quantification of global line-1 repetitive element methylation精制、快速和可再生的高分辨率融化(人力资源管理)的方法适合量化全球第1行重复元素的甲基化
A refined, rapid and reproducible high resolution
melt (HRM)-based method suitable for
quantification of global LINE-1 repetitive element
methylation
Tse et al.
Tse et al. BMC Research Notes 2011, 4:565
/1756-0500/4/565 (28 December 2011)
Tse et al. BMC Research Notes 2011, 4:565
/1756-0500/4/565
TECHNICAL NOTE Open Access
A refined, rapid and reproducible high resolution
melt (HRM)-based method suitable for
quantification of global LINE-1 repetitive
element methylation
1* 2 2 2 3 1*
M Yat Tse , Janet E Ashbury , Nora Zwingerman , Will D King , Sherry AM Taylor and Stephen C Pang
Abstract
Background: The methylation of DNA is recognized as a key mechanism in the regulation of genomic stability
and evidence for its role in the development of cancer is accumulating. LINE-1 methylation status represents a
surrogate measure of genome-wide methylation.
Findings: Using high resolution melt (HRM) curve analysis technology, we have established an in-tube assay that is
linear (r 0.9986) with a high amplification efficiency (90-105%), capable of discriminating between partcipant
samples with small differences in methylation, and suitable for quantifying a wide range of LINE-1 methylation
levels (0-100%)–including the biologically relevant range of 50-90% expected in human DNA. We have optimized
this procedure to perform using 2 μg of starting DNA and 2 ng of bisulfite-converted DNA for each PCR reaction.
Intra- and inter-assay coefficients of variation were 1.44% and 0.49%, respectively, supporting the high
reproducibility and precision of this approach.
Conclusions: In summary,
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