福建农业学报2011，Vol.26Issue(2)：153-158,6.
微生物发酵床垫料微生物总DNA提取方法的研究
Extraction Methods for Bedding Microbial DNA from Fermentation Bed
摘要
Abstract
Traditional plate culture method can not fully reflect the microbial genetic information in microbiological fermentation bed, so obtaining high quality DNA is necessary to study microbial community structure in fermentation bed. In this research we designed and compared six extraction methods for bedding microbial DNA from fermentation bed, mainly in evaluating concentration and purification of extracted DNA. Results indicated that the kit methods have high pertinence, without specific bedding material microorganism DNA extraction kit, the use of excrement, soil and silt kit did not achieve good results. Simply applying SDS or CTAB methods could not effectively eliminate the pollution of humic acid and remove the complex proteins; so that, the ratio of A260 to A280 of the DNA was lower than 1.5 and no result obtained by PCR amplification. Comparatively, in SDS-CTAB method,using 10％ PEG8000 in precipitation obtained the DNA with good integrity and high purity, the concentration of DNA was more than 200 ng·μL-1, the ratio of A260 to A280 was about 1.8, and the PCR amplified targeted the DNA. Results indicated that the SDS-CTAB was an efficient and reliable method for DNA extraction from microbiological fermentation bed and it would be also conducive to the research of molecular ecology.关键词
垫料微生物/DNA提取/SDS-CTAB/16S rDNAKey words
Bedding microorganism/ DNA extraction/ SDS-CTAB/ 16S rDNA分类
生物科学引用本文复制引用
刘云浩,蓝江林,刘波,朱昌雄,郭萍,陈燕萍..微生物发酵床垫料微生物总DNA提取方法的研究[J].福建农业学报,2011,26(2):153-158,6.基金项目
国家水体污染控制与治理科技重大专项(2008ZX07425-002) （2008ZX07425-002）
国家科技支撑计划项目(2008BAD96B07、2009BADC2B02) （2008BAD96B07、2009BADC2B02）
福建省发改委项目(闽发改投资[2008]762号) （闽发改投资[2008]762号）
福建省科技计划--省属公益类科研院所基本科研专项(2010R1020-1) （2010R1020-1）
