A Low-Cost Method of Skin Swabbing for the Collection of DNA Samples from Small Laboratory Fish.
A Low-Cost Method of Skin Swabbing for the Collection of DNA Samples from Small Laboratory Fish.
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DOI：
10.1089/zeb.2016.1348
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发表时间：
2017-02
期刊：
影响因子：
2
通讯作者：
Tilley CA
中科院分区：
文献类型：
--
作者：
Breacker C;Barber I;Norton WH;McDearmid JR;Tilley CA
在麻醉下剪断活鱼的鳍被广泛用于采集DNA提取的样本。另一种可能侵入性较小的替代方法是通过擦拭未麻醉的鱼的皮肤来获取样本。然而，这种方法尚未被广泛应用于生物和生物医学科学的实验室研究中。在这里，我们比较了通过剪鳍和皮肤擦拭技术收集的斑马鱼Danio rerio和三刺刺鱼Gasterbone aculeatus的DNA样本，并测试了一系列DNA提取方法，包括商用试剂盒和成本较低的内部方法。我们验证了聚合酶链式反应分析的方法，并检查了在一起网捕的个体鱼之间交叉污染的潜在风险。我们表明，擦拭可能不需要使用麻醉或止痛剂，提供了一种可靠的替代剪鳍的方法。现在需要进一步的工作，以确定剪鳍和擦拭对鱼类的应激反应和随后的健康的相对影响，从而确定擦拭作为现有DNA采样程序的改进的可能性。
Fin clipping of live fish under anesthesia is widely used to collect samples for DNA extraction. An alternative, potentially less invasive, approach involves obtaining samples by swabbing the skin of nonanesthetized fish. However, this method has yet to be widely adopted for use in laboratory studies in the biological and biomedical sciences. Here, we compare DNA samples from zebrafish Danio rerio and three-spined sticklebacks Gasterosteus aculeatus collected via fin clipping and skin swabbing techniques, and test a range of DNA extraction methods, including commercially available kits and a lower-cost, in-house method. We verify the method for polymerase chain reaction analysis, and examine the potential risk of cross contamination between individual fish that are netted together. We show that swabbing, which may not require the use of anesthesia or analgesics, offers a reliable alternative to fin clipping. Further work is now required to determine the relative effects of fin clipping and swabbing on the stress responses and subsequent health of fish, and hence the potential of swabbing as a refinement to existing DNA sampling procedures.