The effects of preservatives and temperatures on arachnid DNA
The effects of preservatives and temperatures on arachnid DNA
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防腐剂和温度对蜘蛛 DNA 的影响
DOI：
10.1071/is04039
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发表时间：
2005
影响因子：
2.2
通讯作者：
M. Hedin
中科院分区：
文献类型：
--
作者：
C. Vink;Steven M. Thomas;P. Paquin;C. Hayashi;M. Hedin
我们测试了不同防腐剂和温度对可用于 PCR 扩增的蜘蛛和蝎子 DNA 产量的影响。我们的实验旨在模拟六周时间内的现场和实验室条件，测试防腐剂 RNAlater®、丙二醇和各种乙醇浓度。每个保存处理的三个重复在五个不同的温度处理下保存； –80°C、–20°C、2–4°C、19–24°C 和 40°C。提取 DNA 并通过微型凝胶电泳以及高拷贝线粒体 DNA 片段（细胞色素氧化酶亚基 I）和低拷贝核 DNA 片段（肌动蛋白）的 PCR 扩增来评估质量。结果表明，RNAlater® 和丙二醇在高质量 DNA 保存方面明显优于其他防腐剂，并且组织最好在 –80°C 或 –20°C 下保存。如果样本储存在 –20°C 或 –80°C 下，则适合储存在 95% 乙醇中。我们相信，我们的研究结果可以帮助指导生物学家为蜘蛛类、其他节肢动物和无脊椎动物的 DNA 研究选择防腐剂和温度。
We tested the effects of different preservatives and temperatures on the yield of spider and scorpion DNA useable for PCR amplification. Our experiment was designed to simulate conditions in the field and laboratory over a six-week time period, testing the preservatives RNAlater®, propylene glycol, and various ethanol concentrations. Three replicates of each preservation treatment were stored at five different temperature treatments; –80°C, –20°C, 2–4°C, 19–24°C, and 40°C. DNA was extracted and quality was assessed by electrophoresis on mini-gels, and by PCR amplification of high copy mitochondrial DNA fragments (cytochrome oxidase subunit I) and low copy nuclear DNA fragments (actin). Results show that RNAlater® and propylene glycol are significantly better than the other preservatives for high quality DNA preservation and that tissue is best stored at –80°C or –20°C. Storage in 95% ethanol is appropriate if specimens are stored at –20°C or –80°C. We believe our results can help guide biologists in choosing preservatives and temperatures for DNA-based research on arachnids, other arthropods and invertebrates in general.