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比较不同提取方案从鱼鳞片中提取的 DNA 产量。

Comparing DNA yield from fish scales following different extraction protocols.

机构信息

Department of Zoology and Entomology, University of Pretoria, Private Bag X20, Hatfield, 0028, South Africa.

出版信息

Sci Rep. 2022 Feb 18;12(1):2836. doi: 10.1038/s41598-022-06889-y.

DOI:10.1038/s41598-022-06889-y
PMID:35181723
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8857249/
Abstract

Studies on genetic diversity, adaptive potential and fitness of species have become a major tool in conservation biology. These studies require biological material containing a reliable source of DNA which can be extracted and analysed. Recently, non-invasive sampling has become the preferred sampling method of such biological material; particularly when studying endangered species. Elasmoid scales from teleost fish are an example of non-invasive samples from which DNA can successfully be extracted. This study compared different extraction protocols to find an optimal method for extracting DNA from teleost fish scales. This was done with the intent to use the protocol that yielded the highest quantity of DNA on dried, archived scales. The protocols tested in this study included (1) phenol/chloroform with a TNES-urea digestion buffer, (2) phenol/chloroform with an amniocyte digestion buffer and (3) Qiagen DNeasy Blood and Tissue Kit with variations in incubation times and temperatures of each protocol. While the phenol/chloroform with TNES-urea digestion buffer yielded significantly higher concentrations of DNA compared to the other protocols, all protocols followed in this study yielded sufficient quantities of DNA for further downstream applications. Therefore, while there are multiple viable options when selecting a DNA extraction protocol, each research project's individual needs, requirements and resources need to be carefully considered in order to choose the most effective protocol.

摘要

对物种的遗传多样性、适应潜力和适合度的研究已成为保护生物学的主要工具。这些研究需要包含可靠 DNA 来源的生物材料,这些材料可以被提取和分析。最近,非侵入性采样已成为此类生物材料的首选采样方法;特别是在研究濒危物种时。来自硬骨鱼的鲨鱼鳞片就是可以成功提取 DNA 的非侵入性样本的一个例子。本研究比较了不同的提取方案,以找到一种从硬骨鱼鳞片中提取 DNA 的最佳方法。本研究的目的是使用在干燥、存档的鳞片上产生最高 DNA 量的方案。本研究中测试的方案包括 (1) 苯酚/氯仿与 TNES-尿素消化缓冲液,(2) 苯酚/氯仿与羊水消化缓冲液,以及 (3) Qiagen DNeasy Blood 和 Tissue Kit,每个方案都有孵育时间和温度的变化。虽然苯酚/氯仿与 TNES-尿素消化缓冲液产生的 DNA 浓度明显高于其他方案,但本研究中采用的所有方案都产生了足够数量的 DNA,可用于进一步的下游应用。因此,虽然在选择 DNA 提取方案时有多种可行的选择,但需要仔细考虑每个研究项目的个人需求、要求和资源,以选择最有效的方案。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e2e4/8857249/31740062723c/41598_2022_6889_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e2e4/8857249/e5ea3e2ad7c8/41598_2022_6889_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e2e4/8857249/b1de0015b5d8/41598_2022_6889_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e2e4/8857249/1f0852551bae/41598_2022_6889_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e2e4/8857249/706675653d75/41598_2022_6889_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e2e4/8857249/fd9d39418374/41598_2022_6889_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e2e4/8857249/31740062723c/41598_2022_6889_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e2e4/8857249/e5ea3e2ad7c8/41598_2022_6889_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e2e4/8857249/b1de0015b5d8/41598_2022_6889_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e2e4/8857249/1f0852551bae/41598_2022_6889_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e2e4/8857249/706675653d75/41598_2022_6889_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e2e4/8857249/fd9d39418374/41598_2022_6889_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e2e4/8857249/31740062723c/41598_2022_6889_Fig6_HTML.jpg

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