两种标记方法对cDNA芯片杂交信号的初步分析  

Preliminary analysis of cDNA microarray hybridization signal based on two labeling methods

作　　者：杨国淋 赵松[1] 黄小波[1] 马锐[1] 张仙[1] 滑翔 赵玉佳[1] 曹三杰[1] 文翼平[1] 伍锐[1] 文心田[1] 

机构地区：[1]四川农业大学动物医学院动物传染病与基因芯片实验室,四川成都611130

出　　处：《中国兽医科学》2016年第3期338-343,共6页Chinese Veterinary Science

基　　金：公益性行业(农业)科研专项项目(201203056)

摘　　要：为研究两种不同的标记方法对杂交效率的影响,将含NDV的F和HN基因、IBV的M和N基因、ILTV的TK和gB基因的重组质粒作为靶基因,分别采用Cy3直接引物标记法和间接掺入标记法进行PCR扩增。然后将荧光标记的样品与芯片进行杂交,收集扫描后的输出图像和荧光强度信号值,分析两种标记方法对芯片杂交效率的影响。结果显示,Cy3引物标记方法所获得NDV、IBV、ILTV芯片扫描数据结果,图像直观判断均优于间接掺入标记方法。表明Cy3直接引物标记法是一种能够提高杂交的信号值,降低背景信号值,提高杂交效率的方法,有利于检测型基因芯片的应用。In order to research the effect of two different methods on the hybridization efficiency,six of recombinant plasmids as templates which contain the F and HN genes of NDV, M and N genes of IBV,TK and g B genes of ILTV, respectively. And then amplified these target gens with Cy3 direct labeling and indirect incorporation labeling. Hybridization efficiency was analyzed, according to comparing two kinds of methods on the effect of the median,background values and the signal of noise ratio. The result showed that the method of Cy3 direct labeling was better than the indirect labeling according to the scanning parameters and data analysis to the chips of NDV,IBV and ILTV. This study shows that Cy3 direct labeling method which can reduce the background value, so that this method compared to indirect labeling method can improve the efficiency of the hybridization, was beneficial to the application of the detection microarray.

关 键 词：标记方法 杂交信号 基因芯片 

分 类 号：S852.65[农业科学—基础兽医学]