A novel method to quantitate methylation of specific genomic regions.

M Heiskanen, A C Syvänen, H Siitari, S Laine, A Palotie
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引用次数: 10

Abstract

A new solid-phase primer extension method has been developed for the quantitation of methylation differences and is described here. The method is less cumbersome than Southern blot analysis, expresses the results in a numerical format, can be adapted to a microtitration well format, and thus allows the analysis of a large series of samples. The model gene analyzed here is the calcitonin gene, but the method can be adapted to the analysis of methylation alterations in any area of the genome. The primer extension method clearly differentiated hypermethylated samples from normally methylated samples and a range for normal values could be determined. In quantitation experiments the method showed linearity in a range from 2% to 100% malignant blasts diluted with normal leukocytes.

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一种量化特定基因组区域甲基化的新方法。
本文描述了一种新的固相引物延伸法用于甲基化差异的定量。该方法比南方印迹分析更简单,以数值形式表达结果,可以适应微滴定孔格式,因此可以分析大量样品。这里分析的模型基因是降钙素基因,但该方法可以适用于分析基因组任何区域的甲基化改变。引物延伸法明确区分了高甲基化样品和正常甲基化样品,并确定了正常值范围。在定量实验中,该方法在2%到100%的范围内呈线性,恶性细胞被正常白细胞稀释。
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