渔业科学进展2011，Vol.32Issue(2)：117-121,5.
鳗弧菌膜芯片检测条件的优化
Optimization of the detection protocol for membrane chip of Vibrio anguillarum
摘要
Abstract
Gene chip was prepared with the oligonucleotide probes of 6 virulence genes of Vibrio anguillarum dot-applied on a piece of positively charged nylon membrane using the microcaster manual arrayer, followed by cross-linkage under the ultraviolet light. The detection of V. anguillarum virulence genes was performed by hybridizing with the DIG-labeled PCR products on the membrane chip. Here we report the optimization of the concentration of the probe,the time course of hybridization, and the process of washing steps. After optimization, the hybridization time was reduced to 30min, the concentration of the probe as 50μmol/L, and the process of washing was shortened to 3 steps of only 45 min.关键词
基因芯片/尼龙膜/鳗弧菌/毒力基因Key words
Gene chip/ Nylon membrane/ Vibrio anguillarum/ Virulence gene分类
农业科技引用本文复制引用
闫丽,李晨,张庆利,戈蕾,黄倢..鳗弧菌膜芯片检测条件的优化[J].渔业科学进展,2011,32(2):117-121,5.基金项目
国家高技术研究发展计划(863)课题(2006AA100306)和农业公益性行业科研专项经费项目(200803012)共同资助 （863）
