微生物荧光原位杂交(FISH)实验技术  

Microbiology experiment of fluorescence in situ hybridization(FISH) technique

作　　者：陈瑛[1] 任南琪[1] 李永峰[1] 程瑶[1] 

机构地区：[1]哈尔滨工业大学市政环境工程学院

出　　处：《哈尔滨工业大学学报》2008年第4期546-549,575,共5页Journal of Harbin Institute of Technology

基　　金：国家杰出青年科学基金资助项目(50125823);国家重点基础研究发展计划资助项目(G2000026402)

摘　　要：为更好地应用荧光原位杂交(FISH)实验进行微生物群落生态研究工作,对FISH实验原理和影响FISH实验结果准确性的主要因素进行分析和讨论,包括寡核苷酸探针的选择与标记、样品的预处理、杂交液和杂交条件的选择等步骤.总结了一些实验心得,提出关键步骤的操作要点和注意事项,以及FISH技术与其他实验手段相结合的发展前景.建议根据不同检测对象和实验目的,对关键步骤进行改进可以提高实验准确性、简化操作和降低成本.In order to study the ecology of microbial communities by applying Fluorescence in situ hybridization（FISH） technique, this paper analyzed the principle of FISH and the influencing factors of the experimental accuracy. These factors include choosing and marking the oligonucleotide probes, pretreating the samples, and choosing the hybridizing solution and hybridizing conditions. The experimental results were summarized and the main operation points of key procedures were suggested. The future of FISH combining with other techniques was expected. This paper suggests that the increase of the accuracy, the simplification of the procedure and the decrease of the cost can be achieved by improving key procedures according to different samples and researching goals.

关 键 词：微生物生态学 环境微生物 荧光原位杂交 寡核苷酸探针 

分 类 号：Q93[生物学—微生物学]