On-site filtration of water samples for environmental DNA analysis to avoid DNA degradation during transportation
On-site filtration of water samples for environmental DNA analysis to avoid DNA degradation during transportation
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DOI：
10.1007/s11284-016-1400-9
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发表时间：
2016-11-01
影响因子：
2
通讯作者：
Minamoto, Toshifumi
中科院分区：
文献类型：
--
作者：
Yamanaka, Hiroki;Motozawa, Hiromu;Minamoto, Toshifumi
环境DNA （eDNA）分析是确定水生大型生物分布或丰度的一种创新工具。然而，由于eDNA在水中降解迅速，采样和分析之间的长时间延迟可能会阻碍eDNA的量化。在本研究中，我们开发了一种便携式过滤系统，可以现场（和路上）过滤水样。使用室外池塘的水进行(1)现场过滤，(2)冰上运输，(3)环境温度下的水运输，比较6h内eDNA的降解率。第2组和第3组在取样后6 h在实验室过滤。eDNA浓度用实时聚合酶链反应测定两种鱼类线粒体细胞色素b基因的拷贝数。便携式过滤系统具有以下优点：(1)eDNA浓度与采样时保持一致，允许更高的eDNA定量准确性；(2)使用一次性密封塑料袋降低污染风险，并确保在路上过滤；(3)节省时间，因为在采样点之间行驶时可以完成过滤。
Environmental DNA (eDNA) analysis is an innovative tool for determining the distribution or abundance of aquatic macroorganisms. However, because eDNA degrades rapidly in water, long delays between sampling and analysis may hinder eDNA quantification. In the present study, we developed a portable filtration system that enables on-site (and on-the-road) filtration of water samples. Degradation rates of eDNA within 6 h were compared using water from an outdoor pond that was subjected to (1) on-site filtration, (2) transportation of water on ice, and (3) transportation of water at ambient temperature. Groups 2 and 3 were filtered in the laboratory 6 h after sampling. The concentration of eDNA was determined as the copy number of the mitochondrial cytochrome b gene of two fish species using real-time polymerase chain reaction. The portable filtration system offers the following benefits: (1) the eDNA concentration is preserved as is at the time of sampling, permitting higher accuracy of eDNA quantification, (2) use of a disposable sealed plastic bag reduces the risk of contamination and ensures on-the-road filtration, (3) time is saved because filtration can be accomplished when driving between sampling sites.