Accurate quantitation of allele-specific expression patterns by analysis of DNA melting.
Accurate quantitation of allele-specific expression patterns by analysis of DNA melting.
复制标题
通过 DNA 熔解分析准确定量等位基因特异性表达模式。
DOI：
10.1101/gr.6028507
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发表时间：
2007
期刊：
影响因子：
7
通讯作者：
Pfeifer,Karl
中科院分区：
文献类型：
--
作者：
Jeong,Sangkyun;Hahn,Yoonsoo;Rong,Qi;Pfeifer,Karl
表观遗传和遗传机制可以导致二倍体生物体中两个等位基因的表达水平的巨大差异。此外，这些差异可能对个体之间的表型变异至关重要。在这项研究中，我们提出了一种新的程序和算法，以精确和准确地定量每个等位基因的相对表达。该方法利用DNA的差异解链特性，即使在单个碱基对上也不同。通过参考两个纯等位基因的解链特征，可以在数学上解析两个等位基因对任何未知混合物的分数贡献。这些方法具有高度准确性和精密度，因为每个单一解链反应都会产生多个数据点用于分析。最后，我们讨论了这种方法可以更普遍地使用，以准确地定量基因表达相对于已知的标准。
Epigenetic and genetic mechanisms can result in large differences in expression levels of the two alleles in a diploid organism. Furthermore, these differences may be critical to phenotypic variations among individuals. In this study, we present a novel procedure and algorithm to precisely and accurately quantitate the relative expression of each allele. This method uses the differential melting properties of DNAs differing at even a single base pair. By referring to the melting characteristics of the two pure alleles, the fractional contribution of the two alleles to any unknown mixture can be mathematically resolved. These methods are highly accurate and precise because each single melting reaction yields multiple data points for analysis. Finally, we discuss how this approach can be used more generally to accurately quantitate gene expression relative to known standards.