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长度多态性的直接扩增(DALP),或如何在许多物种中获取并鉴定新的遗传标记。

Direct amplification of length polymorphisms (DALP), or how to get and characterize new genetic markers in many species.

作者信息

Desmarais E, Lanneluc I, Lagnel J

机构信息

Laboratoire de Génome et Populations CNRS UPR 9060, Université Montpellier 2 CC. 63, Place E.Bataillon, 34095 Montpellier, France.

出版信息

Nucleic Acids Res. 1998 Mar 15;26(6):1458-65. doi: 10.1093/nar/26.6.1458.

Abstract

Direct amplification of length polymorphisms (DALP) uses an arbitrarily primed PCR (AP-PCR) to produce genomic fingerprints and to enable sequencing of DNA polymorphisms in virtually any species. Oligonucleotide pairs were designed to each produce a specific multi-banded pattern and all the fragments thus generated can be directly sequenced with the same two universal M13 sequencing primers. This strategy combines the advantages of a high resolution fingerprint technique and the possibility of characterizing the polymorphisms. The use of family members as templates in the multi-locus detection step allows a direct test of allele transmission, as well as early mapping of the markers or selection of loci associated with some traits or diseases. We used this method to detect micro-deletions/insertions and microsatellite DNA loci useful in population genetics studies, but it could be applied in many other fields of biology, such as genome mapping for definition of polymorphic sequence tagged sites, directly localized on a genetic map.

摘要

长度多态性直接扩增法(DALP)利用任意引物PCR(AP-PCR)产生基因组指纹图谱,并能够对几乎任何物种的DNA多态性进行测序。设计寡核苷酸对以各自产生特定的多带型模式,并且由此产生的所有片段都可以用相同的两个通用M13测序引物直接测序。该策略结合了高分辨率指纹技术的优点以及表征多态性的可能性。在多位点检测步骤中使用家庭成员作为模板,可以直接测试等位基因传递,以及对标记进行早期定位或选择与某些性状或疾病相关的基因座。我们使用这种方法来检测群体遗传学研究中有用的微缺失/插入和微卫星DNA基因座,但它可应用于生物学的许多其他领域,例如用于定义多态性序列标签位点的基因组图谱绘制,这些位点可直接定位在遗传图谱上。

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