Campanella James J, Smalley John V
Montclair State University, Department of Biology and Molecular Biology, 1 Normal Avenue, Upper Montclair, New Jersey 07043, USA.
BMC Genet. 2006 May 30;7:32. doi: 10.1186/1471-2156-7-32.
The acquisition of high-quality DNA for use in phylogenetic and molecular population genetic studies is a primary concern for evolutionary and genetic researchers. Many non-destructive DNA sampling methods have been developed and are used with a variety of taxa in applications ranging from genetic stock assessment to molecular forensics.
The authors have developed a field sampling method for obtaining high-quality DNA from sunfish (Lepomis) and other freshwater fish that employs a variation on the buccal swab method and results in the collection of DNA suitable for PCR amplification and polymorphism analysis. Additionally, since the circumstances of storage are always a concern for field biologists, the authors have tested the potential storage conditions of swabbed samples and whether those conditions affect DNA extraction and PCR amplification. It was found that samples stored at room temperature in the dark for over 200 days could still yield DNA suitable for PCR amplification and polymorphism detection.
These findings suggest that valuable molecular genetic data may be obtained from tissues that have not been treated or stored under optimal field conditions. Furthermore, it is clear that the lack of adequately low temperatures during transport and long term storage should not be a barrier to anyone wishing to engage in field-based molecular genetic research.
获取用于系统发育和分子群体遗传学研究的高质量DNA是进化和遗传研究人员首要关注的问题。已经开发出许多非破坏性DNA采样方法，并应用于从遗传种群评估到分子法医学等各种分类群。
作者开发了一种从太阳鱼（蓝鳃太阳鱼属）和其他淡水鱼中获取高质量DNA的现场采样方法，该方法采用了颊拭子法的变体，能够收集到适合PCR扩增和多态性分析的DNA。此外，由于储存条件一直是野外生物学家关心的问题，作者测试了拭子样本的潜在储存条件以及这些条件是否会影响DNA提取和PCR扩增。结果发现，在室温黑暗条件下储存超过200天的样本仍能产生适合PCR扩增和多态性检测的DNA。
这些发现表明，可以从未在最佳野外条件下处理或储存的组织中获得有价值的分子遗传数据。此外，很明显，运输和长期储存过程中缺乏足够低的温度不应成为任何希望从事基于野外的分子遗传学研究的人的障碍。
