Ding Chunming, Cantor Charles R
Bioinformatics Program and Center for Advanced Biotechnology, Boston University, 36 Cummington Street, Boston, MA 02215, USA.
Proc Natl Acad Sci U S A. 2003 Jun 24;100(13):7449-53. doi: 10.1073/pnas.1232475100. Epub 2003 Jun 11.
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.
单倍型是几个相位确定的多态性标记的组合,对于疾病关联和染色体进化研究极为重要。在此,我们描述了一种称为M1-PCR的技术(M代表“多重”,1代表“单拷贝DNA分子”),该技术能够对相隔多达24 kb的几个多态性标记进行直接分子单倍型分析。首先将基因组DNA样品稀释至大约单拷贝。通过在同一反应中使用多重PCR和碱基延伸反应对几个多态性标记同时进行基因分型,直接确定单倍型。这种方法不依赖系谱数据,也不需要事先扩增包含所选标记的整个基因组区域。