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历史样本和非侵入性样本:小食蚁兽(Tamandua tetradactyla L.,披毛目)新分离微卫星基因分型错误的一个研究案例

Historical and non-invasive samples: a study case of genotyping errors in newly isolated microsatellites for the lesser anteater (Tamandua tetradactyla L., Pilosa).

作者信息

Clozato Camila L, Moraes-Barros Nadia, Santos Fabrício R, Morgante João S

机构信息

Laboratório de Biologia Evolutiva e Conservação de Vertebrados, Departamento de Genética e Biologia Evolutiva, Instituto de Biociências, Universidade de São Paulo, R. do Matão, 277, 05508-090, São Paulo, Brasil.

出版信息

Mol Ecol Resour. 2014 May;14(3):531-40. doi: 10.1111/1755-0998.12214. Epub 2014 Jan 10.

DOI:10.1111/1755-0998.12214
PMID:24345284
Abstract

Tamandua tetradactyla (Pilosa), the lesser anteater, is a medium-size mammal from South America. Its wide distribution through different landscapes, solitary and nocturnal habits, and the difficulty to capture and contain specimens limit the amount of individuals and populations sampled during fieldworks. These features along with the lack of specific molecular markers for the lesser anteater might be the causes for paucity in population genetic studies for the species. Historical samples from museum specimens, such as skins, and non-invasive samples, such as plucked hair, can be supplementary sources of DNA samples. However, the DNA quantity and quality of these samples may be limiting factors in molecular studies. In this study, we describe nine microsatellite loci for T. tetradactyla and test the amplification success, data reliability and estimate errors on both historical and non-invasive sample sets. We tested nine polymorphic microsatellites and applied the quality index approach to evaluate the relative performance in genotype analysis of 138 historical samples (study skin) and 19 non-invasive samples (plucked hair). The observed results show a much superior DNA quality of non-invasive over historical samples and support the quality index analysis as a practical tool to exclude samples with doubtful performance in genetic studies. We also found a relationship between the age of non-invasive samples and DNA quality, but lack of evidence of this pattern for historical samples.

摘要

小食蚁兽(Tamandua tetradactyla,披毛目)是一种来自南美洲的中等体型哺乳动物。其广泛分布于不同的地貌环境,具有独居和夜行的习性,且捕获和圈养标本存在困难,这些因素限制了野外工作中所采样的个体数量和种群数量。这些特征,再加上缺乏针对小食蚁兽的特定分子标记,可能是该物种种群遗传学研究匮乏的原因。来自博物馆标本(如兽皮)的历史样本以及非侵入性样本(如拔下的毛发)可以作为DNA样本的补充来源。然而,这些样本的DNA数量和质量可能是分子研究中的限制因素。在本研究中,我们描述了9个小食蚁兽的微卫星位点,并测试了历史样本集和非侵入性样本集的扩增成功率、数据可靠性以及估计误差。我们测试了9个多态性微卫星,并应用质量指数方法来评估138个历史样本(研究用兽皮)和19个非侵入性样本(拔下的毛发)在基因型分析中的相对性能。观察结果表明,非侵入性样本的DNA质量远优于历史样本,并支持将质量指数分析作为一种实用工具,以排除在遗传研究中表现存疑的样本。我们还发现了非侵入性样本的年龄与DNA质量之间的关系,但没有证据表明历史样本存在这种模式。

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