快速提取肠道微生物基因组DNA的方法

被引:25
作者
金晶
彭颖
李晓波
机构
[1] 上海交通大学药学院
基金
国家自然科学基金重大研究计划;
关键词
肠道微生物; 简便酚/氯仿抽提法; ERIC-PCR; 16S r DNA-RFLP;
D O I
10.13241/j.cnki.pmb.2007.01.035
中图分类号
Q93-33 [微生物学技术与微生物学实验];
学科分类号
071005 ; 100705 ;
摘要
目的:肠道微生物的研究日益成为热点,如何获取高质量、较完整的肠道菌群基因组DNA是肠道微生物研究中的关键。本文通过对酚/氯仿法提取总DNA过程进行考察和优化,建立一种简便酚/氯仿抽提法。方法:考察和优化酚/氯仿法提取总DNA的过程,并根据DNA产量、纯度以及ERIC-PCR及16S rNDA-RFLP所反映的微生物群落结构特性的指标,并与QIAamp?DNA Stool Mini Kit提取的进行比较,评价了所建立的快速提取方法。结果:用简便酚/氯仿法得到基本完整的基因组DNA,ERIC-PCR和16S rDNA-RFLP结果与QIAamp?DNA Stool Mini Kit法基本相同。结论:该方法快速并成本低,适合肠道微生物研究中总DNA提取,尤其适合处理大批量的样品。
引用
收藏
页码:100 / 103
页数:4
相关论文
共 16 条
[11]  
A simple method for DNA extrac- tion from marine bacteria that produce extracellular materials. LEEYK,KIM HW,LIU CL,et al. Journal of Microbiological Methods . 2003
[12]  
Analysis of the Fecal Microflora of Human Subjects Consuming a Probiotic Prod- uct Containing Lactobacillus rhamnosus DR20. TANNOCK GW,MUNRO K,HARMSEN H J M,et al. Applied and Envi -ronmental Microbiology . 2000
[13]  
Impact of DNA extraction method on bacterial community composition measured by denaturing gradient gel electrophoresis. JULIA RL,ENZINGER C,JOHNSEN K,et al. Soil Biology and Biochem- istry . 2004
[14]  
Intestinal M cells and their role in bacte- rial infection. CLARK MA,MARKark AJ. Int.J.Med.Microbiol . 2003
[15]  
Specificity of enterobacterial repetitive intergenic consensus and repetitive extragenic palindromic polymerase chain reaction for the detection of clonality within the Enterobacter cloacae complex. ANITA N STUMPF,ANDREAS ROGGENKAMP,,HARALD HOFF- MANN. Diagnostic Microbiology and Infectious Disease . 2005
[16]  
A comparison of five methods for extraction of bacterial DNA from human fecal samples. MCORISTAL,JACKSON M,BIRD AR. Journal of Microbiological Methods . 2002