电渗析
异戊醇
色谱法
DNA提取
DNA
乙醇沉淀
化学
环境DNA
限制性酶
萃取(化学)
酒
聚合酶链反应
膜
生物
生物化学
基因
生物多样性
生态学
作者
José Luis Rodríguez‐Mejía,Claudia Martínez‐Anaya,Jorge Luis Folch‐Mallol,Edgar Dantán‐González
标识
DOI:10.1002/elps.200700829
摘要
Extracting DNA from samples of polluted environments using standard methods often results in low yields of poor-quality material unsuited to subsequent manipulation and analysis by molecular biological techniques. Here, we report a novel two-step electrodialysis-based method for the extraction of DNA from environmental samples. This technique permits the rapid and efficient isolation of high-quality DNA based on its acidic nature, and without the requirement for phenol-chloroform-isoamyl alcohol cleanup and ethanol precipitation steps. Subsequent PCR, endonuclease restriction, and cloning reactions were successfully performed utilizing DNA obtained by electrodialysis, whereas some or all of these techniques failed using DNA extracted with two alternative methods. We also show that his technique is applicable to purify DNA from a range of polluted and nonpolluted samples.
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