A method for isolation of soil microbial DNA that is suitable for analysis of microbial cellulase genes
A method for isolation of soil microbial DNA that is suitable for analysis of microbial cellulase genes
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DOI：
10.1080/01496395.2015.1119847
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发表时间：
2016-02
影响因子：
2.8
通讯作者：
Yoshiki Kobayashi;Erika Hayashida;K. Yokoyama;K. Fujii
中科院分区：
文献类型：
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作者：
Yoshiki Kobayashi;Erika Hayashida;K. Yokoyama;K. Fujii
摘要我们建立了一种从土壤中提取纤维素分解微生物DNA的方法，使DNA样品不含土壤有机质(SOM)。该方法包括三个步骤：纤维素分解微生物的繁殖，微生物细胞从SOM中分离，以及用十六烷基三甲基溴化铵(CTAB)提取DNA。用Ficoll-PM400差速离心法成功地将细胞从土壤颗粒和有机质中分离出来。从1g土壤中分离到0.12-0.51kBP的微生物DNA，平均大小为15-20kBP。变性梯度凝胶电泳法显示，从多种纤维素分解菌中提取的DNA得到了回收。
ABSTRACT We developed a method for isolation of DNA of cellulolytic microbies from soil, so that DNA sample was free from soil organic matter (SOM). The method consists of three procedures: propagation of cellulolytic microbes, separation of microbial cells from SOM, and DNA extraction using cetyl trimethyl-ammonium bromide (CTAB). Differential centrifugation with Ficoll-PM400 successfully separated the cells from soil particles and SOM. From 1 g of soil, 0.12-0.51 μg of microbial DNA with an average size of 15-20 kbp was isolated. Denaturing Gradient Gel Electrophoresis showed that DNA from a variety of cellulolytic microbes was recovered.