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A single-nucleotide polymorphism-based approach for rapid and cost-effective genetic wolf monitoring in Europe based on noninvasively collected samples

机译:基于单核苷酸多态性的方法,可基于无创收集的样本在欧洲进行快速且经济高效的遗传狼监测

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Noninvasive genetics based on microsatellite markers has become an indispensable tool for wildlife monitoring and conservation research over the past decades. However, microsatellites have several drawbacks, such as the lack of standardisation between laboratories and high error rates. Here, we propose an alternative single-nucleotide polymorphism (SNP)-based marker system for noninvasively collected samples, which promises to solve these problems. Using nanofluidic SNP genotyping technology (Fluidigm), we genotyped 158 wolf samples (tissue, scats, hairs, urine) for 192 SNP loci selected from the Affymetrix v2 Canine SNP Array. We carefully selected an optimised final set of 96 SNPs (and discarded the worse half), based on assay performance and reliability. We found rates of missing data in this SNP set of <10% and genotyping error of similar to 1%, which improves genotyping accuracy by nearly an order of magnitude when compared to published data for other marker types. Our approach provides a tool for rapid and cost-effective genotyping of noninvasively collected wildlife samples. The ability to standardise genotype scoring combined with low error rates promises to constitute a major technological advancement and could establish SNPs as a standard marker for future wildlife monitoring.
机译:在过去的几十年中,基于微卫星标记的非侵入式遗传学已成为野生动植物监测和保护研究必不可少的工具。但是,微卫星有几个缺点,例如实验室之间缺乏标准化和较高的错误率。在这里,我们提出了一种替代性的基于单核苷酸多态性(SNP)的标记系统,用于无创采集的样本,有望解决这些问题。使用纳流SNP基因分型技术(Fluidigm),我们对158个狼样品(组织,粪便,毛发,尿液)的Affymetrix v2犬SNP阵列中的192个SNP位点进行了基因分型。根据分析的性能和可靠性,我们精心选择了96个SNP的最佳最终组合(并舍弃了最差的一半)。我们发现该SNP集中的数据丢失率低于10%,基因分型错误的发生率接近1%,与其他标记类型的公开数据相比,基因分型准确性提高了近一个数量级。我们的方法提供了一种工具,可以快速,经济高效地对非侵入式采集的野生生物样本进行基因分型。标准化基因型评分和低错误率的能力有望构成一项重大技术进步,并可能将SNP确立为未来野生动植物监测的标准标记。

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