Ethanol and sodium acetate as a preservation method to delay degradation of environmental DNA
Ethanol and sodium acetate as a preservation method to delay degradation of environmental DNA
复制标题
乙醇和乙酸钠作为保存方法可延缓环境 DNA 的降解
DOI：
10.1007/s12686-017-0955-2
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发表时间：
2019
影响因子：
1.1
通讯作者：
J. Amberg
中科院分区：
文献类型：
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作者：
Bridget A. Ladell;L. Walleser;S. McCalla;Richard A. Erickson;J. Amberg
从偏远地区收集的环境DNA（eDNA）样本依赖于DNA的快速稳定。当样品储存在≤ 4 °C时，水样中eDNA的降解最小化。开发一种保存技术，在室温下保持eDNA的完整性，将允许更广泛的位置进行采样。我们评估了乙醇和醋酸钠溶液，以在收集和实验室测试之间保持DNA样本的完整性。对于该评价，将从饲养亚洲鲤鱼的水箱中采集的重复水样置于冰上或保持在室温下。在这两种温度下，水样不处理或用乙醇和乙酸钠溶液（EtOH-NaAc）保存。在采集后的6天内，每天从每种保存方法中取出一部分样本，提取DNA，并采用qPCR测定细胞核和线粒体标志物。结果显示，在没有EtOH-NaAc处理的冰冻样品和保持在室温下的接受EtOH-NaAc处理的样品之间，DNA的持久性相当。我们发现，在室温下保存7天后，使用EtOH-NaAc溶液可以从保存的样品中扩增DNA。
Environmental DNA (eDNA) samples that are collected from remote locations depend on rapid stabilization of the DNA. The degradation of eDNA in water samples is minimized when samples are stored at ≤ 4 °C. Developing a preservation technique to maintain eDNA integrity at room temperature would allow a wider range of locations to be sampled. We evaluated an ethanol and sodium acetate solution to maintain the integrity of the DNA samples for the time between collection and lab testing. For this evaluation, replicate water samples taken from a tank housing Asian carp were placed on ice or held at room temperature. At both temperatures, water samples were left untreated or were preserved with an ethanol and sodium acetate solution (EtOH–NaAc). Every day for 6 days following collection, a subset of the samples was removed from each preservation method and DNA was extracted and nuclear and mitochondrial markers were assayed with qPCR. Results showed comparable persistence of DNA between iced samples without the EtOH–NaAc treatment and samples that received EtOH–NaAc treatment that were kept at room temperature. We found that DNA can be amplified from preserved samples using an EtOH–NaAc solution after up to 7 days at room temperature.
Abe T.：“等翅目。载于：生物多样性百科全书”学术出版社。
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