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评价不同 DNA 提取方法对历史和道路死亡哺乳动物标本的影响。

An evaluation of DNA extraction methods on historical and roadkill mammalian specimen.

机构信息

Centre for Environmental and Climate Science, Lund University, Ecology Building, 223 62, Lund, Sweden.

Department of Biology, Functional Zoology Unit, Lund University, Ecology Building, 223 62, Lund, Sweden.

出版信息

Sci Rep. 2023 Aug 11;13(1):13080. doi: 10.1038/s41598-023-39465-z.

Abstract

Guidelines identifying appropriate DNA extraction methods for both museum and modern biological samples are scarce or non-existent for mammalian species. Yet, obtaining large-scale genetic material collections are vital for conservation and management purposes. In this study, we evaluated five protocols making use of either spin-column, organic solvents, or magnetic bead-based methods for DNA extraction on skin samples from both modern, traffic-killed (n = 10) and museum (n = 10) samples of European hedgehogs, Ericaneus europaeus. We showed that phenol-chloroform or silica column (NucleoSpin Tissue) protocols yielded the highest amount of DNA with satisfactory purity compared with magnetic bead-based protocols, especially for museum samples. Furthermore, extractions using the silica column protocol appeared to produce longer DNA fragments on average than the other methods tested. Our investigation demonstrates that both commercial extraction kits and phenol-chloroform protocol retrieve acceptable DNA concentrations for downstream processes, from degraded remnants of traffic-killed and museum samples of mammalian specimens. Although all the tested methods could be applied depending on the research questions and laboratory conditions, commercial extraction kits may be preferred due to their effectiveness, safety and the higher quality of the DNA extractions.

摘要

对于哺乳动物物种,用于鉴定博物馆和现代生物样本中合适 DNA 提取方法的指南很少或不存在。然而,获取大规模的遗传物质收集对于保护和管理目的至关重要。在这项研究中,我们评估了五种方法,这些方法利用基于离心柱、有机溶剂或磁性珠的方法从现代交通致死(n=10)和博物馆(n=10)的欧洲刺猬(Ericaneus europaeus)皮肤样本中提取 DNA。我们表明,与基于磁性珠的方法相比,苯酚-氯仿或硅柱(NucleoSpin Tissue)方案可获得最高量的 DNA,且纯度令人满意,尤其是对于博物馆样本。此外,与其他测试方法相比,使用硅柱方案进行的提取平均而言似乎产生了更长的 DNA 片段。我们的研究表明,从哺乳动物标本交通致死和博物馆样本的降解残余物中,商业提取试剂盒和苯酚-氯仿方案都可用于下游处理,获得可接受的 DNA 浓度。虽然根据研究问题和实验室条件可以应用所有测试方法,但由于商业提取试剂盒的有效性、安全性和更高质量的 DNA 提取,可能更倾向于使用商业提取试剂盒。

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