Direct molecular haplotyping of long-range genomic DNA with M1-PCR
Direct molecular haplotyping of long-range genomic DNA with M1-PCR
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DOI：
10.1073/pnas.1232475100
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发表时间：
2003-06-24
影响因子：
11.1
通讯作者：
Cantor, CR
中科院分区：
文献类型：
--
作者：
Ding, CM;Cantor, CR
单倍型是几个阶段确定的多态性标记的组合，对于疾病相关性和染色体进化的研究非常有价值。在这里，我们描述了一种称为M1-PCR的技术（M为“多重”和1为“单拷贝DNA分子”），它能够直接分子单倍型的几个多态性标记分离多达24 kb。首先将基因组DNA样品稀释至约单拷贝。单倍型是直接确定的同时基因分型的几个多态性标记在同一反应与多重PCR和碱基延伸反应。该方法不依赖于谱系数据，并且不需要先前扩增含有所选标记的整个基因组区域。
Haplotypes, combinations of several phase-determined polymorphic markers, are extremely valuable for studies of disease association and chromosome evolution. Here we describe a technique called M1-PCR (M for "multiplex" and 1 for "single-copy DNA molecules") that enables direct molecular haplotyping of several polymorphic markers separated by as many as 24 kb. A genomic DNA sample first is diluted to approximately single-copy. The haplotype is directly determined by simultaneously genotyping several polymorphic markers in the same reaction with a multiplex PCR and base extension reaction. This approach does not rely on pedigree data and does not require previous amplification of the entire genomic region containing the selected markers.