淡水总微生物DNA提取方法的比较
Comparison of extraction methods of total microbial DNA from freshwater.
作者信息
Li P, Yang S F, Lv B B, Zhao K, Lin M F, Zhou S, Song X, Tang X M
机构信息
Biotech Research Institute of Shanghai Academy of Agricultural Sciences, Shanghai, China.
Institute of ECO-Environment and Plant Protection, Shanghai Academy of Agricultural Sciences, Shanghai, China.
出版信息
Genet Mol Res. 2015 Jan 30;14(1):730-8. doi: 10.4238/2015.January.30.16.
The demand for molecular analysis of aquatic microbial communities in freshwater has highlighted the need for efficient methods of DNA extraction. The centrifugation method and filtration-membrane method are 2 widely used methods for extracting DNA. The objective of this study was to compare the extraction efficiency of 3 methods, including the centrifugation method, filtration-membrane method, and modified filtration-membrane method, by evaluating the quantity and purity of DNA extracts obtained from water. DNA extraction was analyzed by agarose gel electrophoresis, ultraviolet-spectroscopy, restriction enzyme digestion, and polymerase chain reaction. The results showed that the modified filtration-membrane method was the most efficient for extracting microbial DNA from freshwater with high integrity and purity and is suitable for molecular applications.
对淡水水生微生物群落进行分子分析的需求凸显了高效DNA提取方法的必要性。离心法和滤膜法是两种广泛使用的DNA提取方法。本研究的目的是通过评估从水中获得的DNA提取物的数量和纯度，比较三种方法（包括离心法、滤膜法和改良滤膜法）的提取效率。通过琼脂糖凝胶电泳、紫外光谱、限制性内切酶消化和聚合酶链反应对DNA提取进行分析。结果表明，改良滤膜法从淡水中提取微生物DNA的效率最高，完整性和纯度高，适用于分子应用。
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