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Obtaining high-quality DNA from elusive small mammals using low-tech hair snares

机译:使用技术含量低的圈套器从难以捉摸的小型哺乳动物中获得高质量的DNA

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摘要

Noninvasive sampling approaches are becoming increasingly important for enabling genetic studies of wildlife populations. While a number of methods have been described to noninvasively sample hair from carnivores and medium-sized mammals, they have largely remained untested in elusive small mammals. Here we describe a novel and inexpensive noninvasive hair snare targeted at an elusive small mammal, the American pika (Ochotona princeps). We explore the quality of the sample by assessing PCR amplification success of mitochondrial and nuclear DNA fragments across four commercially available DNA isolation kits and two different quantities of hair in a factorial design. Additionally, we determined the sex of the individual samples using PCR-RFLP of ZFX/ZFY loci. We found that our snare is effective in obtaining hair that yield DNA of sufficient quality and quantity to successfully amplify a range of mitochondrial and nuclear fragment sizes. Specifically, we found the greatest success in amplifying mitochondrial DNA, nuclear microsatellites and ZFX/ZFY loci using at least 25 hairs as starting material and the DNA IQ system. The hair snares thus provide a cost-effective and minimally intrusive approach to sample elusive or rare small mammals. We anticipate that this approach will be useful to obtain samples for molecular studies of the ecology, evolution and conservation of small, elusive mammals.
机译:无损采样方法对于进行野生动植物种群的遗传研究变得越来越重要。虽然已经描述了许多方法来从肉食性动物和中型哺乳动物中无创地取样毛发,但是在可捉摸的小型哺乳动物中,很大程度上仍未进行测试。在这里,我们描述了一种新颖且便宜的无创网罗,该网罗针对一种难以捉摸的小型哺乳动物,美洲小种(美洲短毛猫)。我们通过评估四种设计中的市售DNA分离试剂盒和两种不同量的头发的线粒体和核DNA片段的PCR扩增成功率,来探索样品的质量。此外,我们使用ZFX / ZFY基因座的PCR-RFLP确定了各个样品的性别。我们发现,圈套器可有效获得可产生足够质量和数量的DNA的头发,从而成功扩增一系列线粒体和核片段大小。具体来说,我们发现使用至少25根毛发作为起始材料和DNA IQ系统,在扩增线粒体DNA,核微卫星和ZFX / ZFY基因座方面取得了最大的成功。因此,毛圈套圈提供了一种成本低廉且侵入性最小的方法来采样难以捉摸的或稀有的小型哺乳动物。我们预计,这种方法将有助于获得用于小型,难以捉摸的哺乳动物的生态学,进化和保护的分子研究的样本。

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