Single-molecule dilution and multiple displacement amplification for molecular haplotyping

被引：18

作者：

Paul, P ^{[1
]}

Apgar, J ^{[1
]}

机构：

[1] Cleveland Clin, Allogen Labs, Cleveland, OH 44106 USA

来源：

BIOTECHNIQUES | 2005年 / 38卷 / 04期

关键词：

D O I：

10.2144/05384ST01

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Separate haploid analysis is frequently required for heterozygous genotyping to resolve phase ambiguity or confirm allelic sequence. We demonstrate a technique of single-molecule dilution followed by multiple strand displacement amplification to haplotype polymorphic alleles. Dilution of DNA to haploid equivalency, or a single molecule, is a simple method for separating di-allelic DNA. Strand displacement amplification is a robust method for nonspecific DNA expansion that employs random hexamers and phage polymerase Phi29 for double-stranded DNA displacement and primer extension, resulting in high processivity and exceptional product length. Single-molecule dilution was followed by strand displacement amplification to expand separated alleles to microgram quantities of DNA for more efficient haplotype analysis of heterozygous genes.

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收藏

页码：553 / +

页数：5

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