摘要
通过改进裂解温度和延长裂解时间并增加苯酚/氯仿洗脱次数的DNA提取方法获得南京玄武湖底泥中的DNA,通过PCR法来扩增微囊藻的16SrRNA基因.结果表明在所有采样点中均得到微囊藻基因组DNA,并且纯度较高,OD_(260)/ OD_(280)均高于1.54,最高值达到1.89.PCR的扩增结果显示所有样点的DNA都得到212 bp大小的微囊藻16SrRNA基因片断,表明这种方法可以有效的从底泥中提取微囊藻的DNA,从而为研究底泥微囊藻生理生态及其越冬、上浮、形成水华的机理提供更有利的方法.
A method for DNA extraction from sediment samples of Lake Xuanwu, Nanjing, China was developed, and purification and efficience of extracted DNAs were evaluated through ultraviolet spectrum OD260/OD2s0 ratios and PCR amplification of 16S rDNA specific for Microcystis. The results showed that sediment samples from all sites in the study obtained quite purified genomic DNAs with OD260/OD280 ratios from 1.54 to 1.89, and Microcystis 16S rRNA gene fragments have been detected in all sampling sites, which indicating this method can be used for molecular detection of Microcystis cells in sediments.
出处
《湖泊科学》
EI
CAS
CSCD
北大核心
2007年第4期492-496,共5页
Journal of Lake Sciences
基金
中国科学院领域前沿(055102-1-501)
国家"863"重大科技项目(2003AA601100-2)
国家"十五""211工程"重大项目"不同时空尺度环境演变和生态建设"
江苏省重点科技专项(BM2002701)联合资助.
关键词
底泥
微囊藻
DNA
水华
蓝藻
Sediment, Microcystis, DNA, waterbloom, cyanobacteria