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Denaturing gradient gel electrophoresis를 이용한 한국의 논 토양 미생물 다양성 분석 방법
최명은 ( Myeongeun Choe ),홍성준 ( Sung Jun Hong ),임종희 ( Jong Hui Lim ),곽윤영 ( Yun Young Kwak ),백창기 ( Chang Gi Back ),정희영 ( Hee Young Jung ),이인중 ( In Jung Lee ),신재호 ( Jae Ho Shin ) 한국응용생명화학회 2013 Journal of Applied Biological Chemistry (J. Appl. Vol.56 No.2
Soil microbes are important integral components of soil ecosystem which have significant and diverse role in organic matter decomposition, nitrogen cycling, and nitrogen fixation. In this study an effective denaturing gradient gel electrophoresis (DGGE) method was employed for paddy soil microbial diversity survey. For optimum paddy soil microbial DNA extraction, different methods such as Lysis buffer, skim milk bead, sodium phosphate buffer, Epicentre Soil Master DNA extraction kit (Epicentre, USA) and Mo Bio Power Soil DNA kit (MO BIO, USA) methods were utilized. Among all the method, using Mo Bio Power Soil kit was most effective. DGGE analysis of Bacteria was carried out at 6% polyacylamide gel and 45-60% denaturing gradient in the optimal conditions. Whereas DGGE analysis of fungi was done at 6% polyacrylamide gel and 45-80% denaturing gradient in the optimal conditions. By applying the above assay, it was found that variation within the microbial community of paddy soil occurs by a factor of time. DGGE say used in this study through for a variety of soil microbial analysis suggests the potential use of this method.