The room temperature preservation of filtered environmental DNA samples and assimilation into a phenol-chloroform-isoamyl alcohol DNA extraction.
The room temperature preservation of filtered environmental DNA samples and assimilation into a phenol-chloroform-isoamyl alcohol DNA extraction.
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过滤的环境DNA样品的室温保存，并同化为酚类氯仿 - 异氧氨基醇DNA提取。
DOI：
10.1111/1755-0998.12281
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发表时间：
2015-01
影响因子：
7.7
通讯作者：
Lodge DM
中科院分区：
文献类型：
--
作者：
Renshaw MA;Olds BP;Jerde CL;McVeigh MM;Lodge DM
目前针对过滤后的大型微生物环境DNA（eDNA）的研究通常依赖于各个阶段的冷环境温度，包括将水样从现场运输到实验室以及将水和/或过滤后的样品储存在实验室中。这对在冷藏和冷冻储存困难或样品必须长途运输以进行进一步处理和筛选的地点进行实地采集造成了实际限制。该研究证明了eDNA在室温（20 °C）下在两种裂解缓冲液（CTAB和Longmire’s）中成功保存超过2周的时间。此外，保存的eDNA样品被无缝整合到苯酚-氯仿-异戊醇（PCI）DNA提取方案中。eDNA提取的成功应用于多种类型的滤膜表明，这里评估的方法可以广泛应用于未来的eDNA研究。我们的研究结果还表明，对于许多种类的研究最近报道的大型微生物eDNA，检测概率可以增加，并在较低的成本，通过利用Longmire的保存缓冲液与PCI DNA提取。
Current research targeting filtered macrobial environmental DNA (eDNA) often relies upon cold ambient temperatures at various stages, including the transport of water samples from the field to the laboratory and the storage of water and/or filtered samples in the laboratory. This poses practical limitations for field collections in locations where refrigeration and frozen storage is difficult or where samples must be transported long distances for further processing and screening. This study demonstrates the successful preservation of eDNA at room temperature (20 °C) in two lysis buffers, CTAB and Longmire's, over a 2-week period of time. Moreover, the preserved eDNA samples were seamlessly integrated into a phenol–chloroform–isoamyl alcohol (PCI) DNA extraction protocol. The successful application of the eDNA extraction to multiple filter membrane types suggests the methods evaluated here may be broadly applied in future eDNA research. Our results also suggest that for many kinds of studies recently reported on macrobial eDNA, detection probabilities could have been increased, and at a lower cost, by utilizing the Longmire's preservation buffer with a PCI DNA extraction.
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