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序列标签微卫星分析(STMP):改进侧翼目标简单序列重复(SSR)的DNA序列分离方法

Sequence tagged microsatellite profiling (STMP): improved isolation of DNA sequence flanking target SSRs.

作者信息

Hayden M J, Good G, Sharp P J

机构信息

Plant Breeding Institute, University of Sydney, PMB 11, Camden, NSW 2570, Australia.

出版信息

Nucleic Acids Res. 2002 Dec 1;30(23):e129. doi: 10.1093/nar/gnf129.

Abstract

Sequence tagged microsatellite profiling (STMP) enables the rapid development of large numbers of co-dominant DNA markers, known as sequence tagged microsatellites (STMs). Each STM is amplified by PCR using a single primer specific to the conserved DNA sequence flanking the microsatellite repeat in combination with a universal primer that anchors to the 5'-ends of the microsatellites. It is also possible to convert STMs into conventional microsatellite, or simple sequence repeat (SSR), markers that are amplified using a pair of primers flanking the repeat sequence. Here, we describe a modification of the STMP procedure to significantly improve the capacity to convert STMs into conventional SSRs and, therefore, facilitate the development of highly specific DNA markers for purposes such as marker-assisted breeding. The usefulness of this technique was demonstrated in bread wheat.

摘要

序列标签微卫星分析(STMP)能够快速开发大量共显性DNA标记,即序列标签微卫星(STM)。每个STM通过PCR扩增,使用一个特异于微卫星重复序列侧翼保守DNA序列的单一引物,结合一个锚定在微卫星5'端的通用引物。也可以将STM转化为传统的微卫星或简单序列重复(SSR)标记,使用一对位于重复序列侧翼的引物进行扩增。在这里,我们描述了对STMP程序的一种改进,以显著提高将STM转化为传统SSR的能力,从而便于开发用于标记辅助育种等目的的高度特异性DNA标记。该技术的实用性在面包小麦中得到了证明。

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