摘要
从常温研磨、蛋白酶K和R nase的加入对传统CTAB法进行改良,并结合试剂盒法,提出了一种高效提取高温大曲微生物总DNA的方法。该方法获得的总基因片段大小约21Kb;A260/A280=1.878;A260/A230=1.706;PCR反应抑制物少,可直接用于16S rDNA的扩增;并通过构建克隆文库技术和变性梯度凝胶电泳(DGGE)分析后发现高温大曲中细菌多样性丰富,能一定程度上反映微生物群落结构和组成。研究结果表明该方法提取的DNA适用于芝麻香高温大曲中微生物的分子生态学研究。
This paper conducted a research on the methods of DNA extraction and defined an mneliorative method. The obtained total genome fragment extracted with this method was about 21kb in size and the purity was A260/A280=1.878, A260/A230=1.706, and can be directly applied to PCR amplification of 16S rDNA. With further validation by cloning library and DGGE, the DNA can represent the microbe community. These resuhs indicate that the total DNA extracted with this method applies to the study on the molecular biology of high temperature Daqu.
出处
《酿酒》
CAS
2012年第4期33-37,共5页
Liquor Making
基金
国家科技支撑计划课题(2012BAK17B11)
