摘要： 应用马铃薯晚疫病水平抗性相关的1 009条表达序列标签(ESTs)所制备的cDNA微阵列和荧光信号Cy3,Cy5杂交系统,通过同一样品RNA的自身比较试验及与不同样品RNA的差异分析试验,对该技术的重复性和可靠性进行了验证分析。结果表明,在自身比较试验中,芯片杂交的假阳性率仅为0.79%,相关系数高达0.981 1;在差异分析试验中,同向标记、正反向标记、同一芯片内2个重复杂交结果的相关系数分别为0.966 0,0.889 5,0.980 2。研究结果证实,cDNA芯片技术具有高度的可靠性,可以鉴定出显著差异表达基因,并构建高质量的基因表达数据库,通过3次生物学重复和2次技术重复试验(采用正、反向标记)即可克服绝大部分试验误差。

关键词: cDNA微阵列, 差异表达基因, 可靠性

Abstract: Self-self comparison experiment of same samples and differential analysis experiments of different samples were performed to verify the reproducibility and reliability of this technology using cDNA microarray containing 1 009 expressed sequence tags(ESTs)associated with quantitative resistance to P.infestans and Cy3/Cy5 hybridization system.The results indicated that the false positive rate was 0.79%,and the correlation coefficient was high to 0.9811 in self-self comparison experiment.In differential analysis experiments,the correlation coefficient of the replications between same dye-labelled slides was 0.966 0,whereas that of the replications between dye-swapped slides was 0.889 5 and that on the same slide was 0.980 2.Our results demonstrated the high reproducibility and reliability of cDNA microarray hybridization.Based on this technology,the significantly differentially expressed genes could be identified,and the high-quality databank of gene expression may be constructed.The most of the experiment errors could be reduced by three biological replicates and two technological replicates(dye-swapped experiments).

Key words: cDNA microarray, Differentially expressed genes, Reliability

中图分类号: 

• S532.01