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用于细菌群落PCR-DGGE分析的根际土壤样品中DNA的提取与纯化。

Extraction and purification of DNA in rhizosphere soil samples for PCR-DGGE analysis of bacterial consortia.

作者信息

Maarit Niemi R, Heiskanen I, Wallenius K, Lindström K

机构信息

Finnish Environment Institute, PO Box 140 FIN-00251, Helsinki, Finland.

出版信息

J Microbiol Methods. 2001 Jul;45(3):155-65. doi: 10.1016/s0167-7012(01)00253-6.

Abstract

Application of DNA fingerprinting methods enables the detection of diverse members of soil bacterial consortia, even including those bacteria not yet cultivated. However, extraction and purification of DNA from soil samples without bias is difficult. We compared five different DNA isolation methods and three purification methods for rhizosphere soil samples. Purified DNA extracts were amplified in PCR using universal bacterial primers and the PCR products were analysed with denaturing gradient gel electrophoresis (DGGE) for the visualisation of DNA bands representing dominant bacterial species. Both the isolation and purification methods affected the apparent bacterial community structure of the samples.

摘要

应用DNA指纹图谱方法能够检测土壤细菌群落中的各种成员,甚至包括那些尚未培养的细菌。然而,从土壤样品中无偏差地提取和纯化DNA是困难的。我们比较了五种不同的DNA提取方法和三种用于根际土壤样品的纯化方法。使用通用细菌引物在PCR中扩增纯化的DNA提取物,并使用变性梯度凝胶电泳(DGGE)分析PCR产物,以可视化代表优势细菌种类的DNA条带。提取和纯化方法都影响了样品中明显的细菌群落结构。

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