摘要
目的研究采用荧光标记的通用引物扩增或在扩增中掺入荧光标记dNTP两种限制性荧光标记方法标记探针对提高表达谱基因芯片杂交结果信噪比的作用。方法酵母mRNA经常规逆转录标记方法及两种限制性标记方法进行标记,探针纯化后与酵母表达谱基因芯片进行杂交,在同等条件下进行杂交后清洗和芯片扫描检测。结果两种限制性标记方法与直接逆转录标记相比杂交结果背景低,信噪比及灵敏度高,其中以荧光标记的通用引物扩增效果最佳。结论采用限制性标记方法后杂交结果信噪比提高,有利于基因芯片技术的应用。
Objective To study the signal-to-noise ratio (SNR) of two restricted fluorescence labeling methods for examining gene expression profile by microarray hybridization. Method Samples of Saccharomyces cerevisiae mRNA was labeled by traditional reverse transcription method and 2 restriction fluorescent labeling methods using respectively Cy-universal primer and extension incorporated Cy-dNTP. The labeled samples were examined by the microarray, followed by washing and scanning under the same conditions. Results The two restriction labeling methods showed superior results with lowered background and enhanced SNR and sensitivity, and Cy-universal primer labeling presented the best results. Conclusion SNR can be enhanced by the restriction labeling methods, which improve the applicability of microarray technology.
出处
《第一军医大学学报》
CSCD
北大核心
2003年第2期124-126,共3页
Journal of First Military Medical University
基金
国家自然科学基金(39880032)
广州市重点科技攻关项目(99202201)~~
关键词
基因芯片技术
荧光标记
酵母
杂交
信噪比
restriction labeling
fluorescent labeling
microarray
restriction display polymerase chain reaction
