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为普通宽吻海豚(Tursiops truncatus)开发基因组资源:153 个单核苷酸多态性和 53 个基因分型检测的分离和特征描述。

Developing genomic resources for the common bottlenose dolphin (Tursiops truncatus): isolation and characterization of 153 single nucleotide polymorphisms and 53 genotyping assays.

机构信息

Department of Biology, University of Louisiana at Lafayette, Lafayette, LA 70504, USA.

出版信息

Mol Ecol Resour. 2012 Nov;12(6):1124-32. doi: 10.1111/1755-0998.12008. Epub 2012 Sep 14.

DOI:10.1111/1755-0998.12008
PMID:22978635
Abstract

Although single nucleotide polymorphisms (SNPs) are commonly used in human genetics, they have only recently been incorporated into genetic studies of non-model organisms, including cetaceans. SNPs have several advantages over other molecular markers for studies of population genetics: they are quicker and more straightforward to score, cross-laboratory comparisons of data are less complicated, and they can be used successfully with low-quality DNA. We screened portions of the genome of one of the most abundant cetaceans in U.S. waters, the common bottlenose dolphin (Tursiops truncatus), and identified 153 SNPs resulting in an overall average of one SNP every 463 base pairs. Custom TaqMan(®) Assays were designed for 53 of these SNPs, and their performance was tested by genotyping a set of bottlenose dolphin samples, including some with low-quality DNA. We found that in 19% of the loci examined, the minor allele frequency (MAF) estimated during initial SNP ascertainment using a DNA pool of 10 individuals differed significantly from the final MAF after genotyping over 100 individuals, suggesting caution when making inferences about MAF values based on small data sets. For two assays, we also characterized the basis for unusual clustering patterns to determine whether their data could still be utilized for further genetic studies. Overall results support the use of these SNPs for accurate analysis of both poor and good-quality DNA. We report the first SNP markers and genotyping assays for use in population and conservation genetic studies of bottlenose dolphins.

摘要

虽然单核苷酸多态性 (SNP) 在人类遗传学中被广泛应用,但直到最近才被纳入非模式生物(包括鲸目动物)的遗传研究中。SNP 相对于其他分子标记在群体遗传学研究中有几个优势:它们更容易评分,实验室间数据比较更简单,并且可以成功用于低质量 DNA。我们筛选了美国水域中最丰富的鲸目动物之一——宽吻海豚 (Tursiops truncatus) 的基因组部分,并鉴定出 153 个 SNP,平均每 463 个碱基对就有一个 SNP。为其中 53 个 SNP 设计了定制的 TaqMan(®) 检测,并用一组宽吻海豚样本对其性能进行了测试,其中一些样本的 DNA 质量较低。我们发现,在 19%的检测基因座中,使用包含 10 个个体的 DNA 池进行初始 SNP 鉴定时估计的次要等位基因频率 (MAF) 与对 100 多个个体进行基因分型后的最终 MAF 有显著差异,这表明在基于小数据集推断 MAF 值时需要谨慎。对于两个检测,我们还对异常聚类模式的基础进行了特征描述,以确定其数据是否仍可用于进一步的遗传研究。总体结果支持使用这些 SNP 进行准确分析,无论 DNA 质量好坏。我们报告了第一个 SNP 标记和用于宽吻海豚种群和保护遗传学研究的基因分型检测。

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