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Investigating a simplified method for noninvasive genetic sampling in East African mammals using silica dried scat swabs

机译:研究一种使用二氧化硅干燥的粪便拭子在东非哺乳动物中进行非侵入性基因采样的简化方法

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摘要

Swabbing scat has proved to be an effective noninvasive method to collect DNA from mammals in the field. Previously, this method has relied on preservative liquids or freezing to preserve the DNA collected on swabs. In this study, we determine the effectiveness of using silica to simply dry the swab in field as an alternative way to prevent DNA degredation. Four species were included in the study; reticulated giraffe, impala, fringe‐eared oryx, and lion. Swabs were taken at multiple time points for giraffe and impala scat samples, with the lion and oryx sampled opportunistically. Mitochondrial DNA was successfully amplified and sequenced from scat swabs from all species; however, effectiveness varied between species, with 81.8% amplification success rate from swabs taken from impala scat compared to 25% amplification success rate in giraffe. This variation in success rate was overcome by taking multiple swabs, thus increasing the probability of a successful amplification. The true merit of this method is in its simplicity and cheapness; no preservative liquids were required to be brought into the field, at no stage in the 2 weeks of field sampling were samples frozen, and no commercial kits were used for DNA extraction.
机译:事实证明,拭粪是一种有效的非侵入性方法,可从野外哺乳动物收集DNA。以前,此方法依靠防腐液或冷冻来保存拭子上收集的DNA。在这项研究中,我们确定了使用二氧化硅在野外简单干燥棉签作为防止DNA降解的另一种方法的有效性。研究中包括四种。网状长颈鹿,黑斑羚,流苏的羚羊和狮子。在多个时间点对长颈鹿和黑斑羚粪便样本进行拭子采集,同时对狮子和大羚羊进行了机会性采样。从所有物种的粪便拭子中成功地扩增并测序了线粒体DNA。但是,不同物种之间的效果有所不同,从黑斑羚粪便中抽取的拭子的扩增成功率为81.8%,而长颈鹿的扩增成功率为25%。通过多次拭子可以克服成功率的这种变化,从而增加成功扩增的可能性。这种方法的真正优点在于它的简单性和廉价性。无需将防腐剂液体带入田间,在田间采样的两周内任何阶段均无需冷冻样品,也无需使用商业试剂盒进行DNA提取。

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