一种河口沉积物总DNA的高效提取方法  

An Efficient Method for DNA Extraction from Estuarine Sediments Microorganism

作　　者：王东升[1] 黄江丽[1] 谭胤静[1] 田晓娟[1] 张志红[1] 丁建南[1] 

机构地区：[1]江西省科学院生物资源研究所,南昌330096

出　　处：《江西科学》2014年第3期323-327,共5页Jiangxi Science

基　　金：国家自然科学基金项目(31260556);江西省科技支撑计划项目(2013BBF60076);江西省自然科学基金(2010BNA09100);江西省科学院博士基金项目(2011YYB05)

摘　　要：研究主要目的是从河口沉积物中提取高质量DNA,为后期应用分子生物学技术研究河口沉积物微生物功能基因及种群分析等奠定基础。用CTAB-SDS-冻融法,样品预处理-CTAB-SDS-冻融法,样品预处理-CTABSDS-冻融-DNA重沉淀法和土壤总DNA提取试剂盒法提取3种河口沉积物总DNA,通过DNA琼脂糖凝胶电泳、DNA纯度和浓度及16S rDNA的PCR扩增分别对这4种方法进行评价。结果显示,样品预处理-CTABSDS-冻融法-DNA重沉淀所提取的DNA质量最高,可以用于后续的分子生物学研究。The main purpose of this research was to extract high quality DNA from estuarine sediments for studying microbial functional gene and population analysis by molecule biology technology. We extracted the total DNA from 3 kinds of estuarine sediments with CTAB-SDS-frozen-thawing- method, sample pretreatment-CTAB-SDS-frozen-thawing-method, sample pretreatment-CTAB-SDS- frozen-thawing-precipitate DNA-method and soil total DNA Extraction Kit, and assessed these DNA extraction methods by DNA agarose gel electrophoresis, DNA purity and concentration as well as PCR amplification 16S rDNA gene. The results showed that extracted total DNA by sample pretreatment-CTAB-SDS-frozen-thawing-preeipitate DNA-method had the highest quality and could be used for subsequent molecule biology studies.

关 键 词：河口沉积物 环境微生物 DNA提取和纯化 方法优化 

分 类 号：Q939.94[生物学—微生物学]