An optmized protocol for simultaneous extraction of DNA and RNA from soils

Brazilian Journal of Microbiology

Rodrigo Costa¹  Newton C.m. Gomes¹  Annett Milling¹  Kornelia Smalla¹ 

[1],Federal Biological Research Centre for Agriculture and Forestry (BBA),Germany

关键词: RNA extraction;    nucleic acids;    rhizosphere;    microbial communities;    extração de RNA;    ácidos nucléicos;    rizosfera;    comunidades microbianas;   

DOI  :  10.1590/S1517-83822004000200011

来源: SciELO

【 摘 要 】

In this work we report an optimized protocol for simultaneous extraction of DNA and RNA from soil matrices. Treatment of soil matrices with ethanol followed by bead-beating worked as a successful strategy to lyse the cells without considerable degradation of nucleic acids, resulting in DNA and RNA of good yield and integrity. The reverse transcribed RNA could be amplified with primers targeting a glutamine synthetase (glnA) gene fragment. From both DNA and cDNA, 16S rDNA fragments were amplified and analyzed by Denaturing Gradient Gel Electrophoresis (DGGE). The method was applied to soil and rhizosphere (strawberry and oilseed rape) samples. Two other protocols for the extraction of nucleic acids from soil were applied to the same set of samples in order to compare the methods in terms of efficiency and reproducibility. The DGGE profiles indicated no relevant differences between the patterns obtained. The method described here is suitable for rapid processing of many samples and therefore appropriate for ecological studies.

【 授权许可】

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