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从感兴趣的AFLP位点轻松获取核苷酸序列的两种方法。

Two methods to easily obtain nucleotide sequences from AFLP loci of interest.

作者信息

Paris Margot, Meyer Claire-Lise, Blassiau Christelle, Coissac Eric, Taberlet Pierre, Després Laurence

机构信息

Institute of Integrative Biology, ETH Zurich, Zurich, Switzerland.

出版信息

Methods Mol Biol. 2012;888:91-108. doi: 10.1007/978-1-61779-870-2_6.

Abstract

Genome scans based on anonymous Amplified Fragment Length Polymorphism (AFLP) markers scattered throughout the genome are becoming an increasingly popular approach to study the genetic basis of adaptation and speciation in natural populations. A shortcoming of this approach is that despite its efficiency to detect signatures of selection, it can hardly help pinpoint the specific genomic region(s), gene(s), or mutation(s) targeted by selection. Here, we present two methods to be undertaken after performing an AFLP-based genome scan to easily obtain the sequences of AFLP loci detected as outliers by population genomics approaches. The first one is based on the gel excision of the target AFLP fragment, after simplification of the AFLP fingerprint and separation of the fragments by migration. The second one is a combination of classical AFLP protocol and 454 pyrosequencing.

摘要

基于散布于整个基因组的匿名扩增片段长度多态性(AFLP)标记进行的基因组扫描,正日益成为研究自然种群适应和物种形成遗传基础的一种流行方法。该方法的一个缺点是,尽管它能有效检测选择信号,但几乎无法帮助确定选择所针对的特定基因组区域、基因或突变。在此,我们提出两种在基于AFLP的基因组扫描后采用的方法,以便轻松获得通过群体基因组学方法检测为异常值的AFLP位点的序列。第一种方法是在简化AFLP指纹并通过迁移分离片段后,对目标AFLP片段进行凝胶切除。第二种方法是经典AFLP方案与454焦磷酸测序的结合。

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