首页> 外文期刊>European journal of wildlife research >The effect of sample age and storage method on DNA yield and microsatellite amplification from baboon (Papio ursinus) faecal samples
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The effect of sample age and storage method on DNA yield and microsatellite amplification from baboon (Papio ursinus) faecal samples

机译:样本年龄和保存方法对狒狒粪便样本DNA产量和微卫星扩增的影响

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摘要

Sampling methods that allow DNA collection without the physical handling of animals are popular in conservation genetics, but DNA isolated from faecal samples may be degraded, potentially leading to erroneous microsatellite genotyping results. We collected baboon faecal samples from fresh to 1-week post-defecation in a controlled sampling environment and preserved these using three storage techniques. After DNA isolation and quantification, the samples were genotyped at eight microsatellite loci. Quantitatively, DNA yield was highest when using silica as storage medium. However, microsatellite amplification from samples stored in 95% ETOH were most successful, with 100% success for fresh samples and dropping only marginally to 87.5% of loci for samples collected after 1 week. This disparity between quantitative and qualitative results suggests that total DNA concentrations do not necessarily provide a reliable indication of the amount of target DNA present in a DNA isolate. Our results nevertheless confirm that microsatellite fragments can successfully be amplified from faecal samples up to 1 week post-defecation, with careful selection of storage protocols and loci.
机译:在保护遗传学中,采样方法允许不进行动物的物理处理而收集DNA,这在保护遗传学中很普遍,但是从粪便样品中分离的DNA可能会降解,从而可能导致错误的微卫星基因分型结果。我们在受控的采样环境中收集了从新鲜到粪便后1周的狒狒粪便样本,并使用三种存储技术对其进行了保存。 DNA分离和定量后,在八个微卫星基因座对样品进行基因分型。从数量上讲,使用二氧化硅作为存储介质时,DNA产量最高。但是,从存储在95%ETOH中的样品进行的微卫星扩增最为成功,新鲜样品的成功率为100%,1周后收集的样品仅略微下降到基因座的87.5%。定量和定性结果之间的这种差异表明总DNA浓度不一定能可靠地表明DNA分离物中存在的目标DNA的量。然而,我们的结果证实,通过仔细选择存储方案和基因座,可以在粪便后1周内成功地从粪便样品中扩增出微卫星片段。

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