An efficient and safe method for the extraction of total DNA from shed frog skin
An efficient and safe method for the extraction of total DNA from shed frog skin
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DOI：
10.1007/s12686-019-01104-z
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发表时间：
2020-06-01
影响因子：
1.1
通讯作者：
Bai, Xiujuan
中科院分区：
文献类型：
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作者：
Xu, Yuan;Guan, Tianming;Bai, Xiujuan
从组织中分离高质量DNA的有效方案对于分子生物学的许多方面都是必不可少的。在涉及青蛙的分子研究中，蜕皮作为高质量DNA的来源经常被忽视。本研究采用酚-氯仿、DNeasy提取试剂盒和改良的盐析法从青蛙蜕皮中提取DNA，并与骨骼肌和脚趾的DNA进行比较。我们发现，改进的盐析程序获得了最高的DNA产量，并避免使用有毒的化学物质，如苯酚，氯仿，异戊醇，和/或异硫氰酸胍。东北林蛙蜕皮组织DNA含量较低。然而，从脱落的皮肤DNA是有用的mtDNA和核DNA的扩增使用PCR，是林蛙和蟾蜍gargarizans。这些结果表明，改良盐析法提取蛙皮DNA是一种安全有效的方法，可用于蛙的保护遗传学研究和群体遗传学研究。
An efficient protocol for the isolation of high-quality DNA from tissues is essential to many aspects of molecular biology. Shed skin is often overlook as a source for high-quality DNA in molecular studies involving frog. In this study, DNA from shed skin of frog was extracted using phenol-chloroform, the DNeasy extraction kit (Qiagen) and an improved salting-out protocol, in comparison with those of skeletal muscle and toe. We found that the improved salting-out procedure obtained the highest DNA yield, and avoided using toxic chemicals such as phenol, chloroform, isoamyl alcohol, and/or guanidine isothiocyanate. The quantity of DNA from shed skin was lower compared to other tissue in Rana dybowskii. Nevertheless, DNA from shed skin was useful for amplifing mtDNA and nuclear DNA using PCR, as were Rana amurensis and Bufo gargarizans. These results indicate that extrating DNA from shed frog skin by improved salting-out protocol is an efficient and safe method for the conservation genetic studies and population genetics without destroy in frogs.