Evaluation of Two Different Sample Labeling Methods on Background Signal Intensities for 60 mer Oligonucleotide Microarrays
两种标记方法对60mer寡核苷酸芯片背景信号影响的初步分析

The effects of two different sample labeling methods on background signal intensities for high-density 60mer oligonucleotide microarray were investigated. Peripheral blood samples from five disease and five control subjects were collected. Total RNA targets from peripheral blood mononuclear cells were extracted and labeled with RD-PCR protocol, which were hybridized to Agilent Human 1B oligonucleotide microarrays in a two-color comparative format. The positive control targets were labeled with the directly incorporated fluorescently-labeled dNTP labeling. The SPSS program was performed to test normality of the dataset, variance homogeneity between the groups, coefficients of variation (CV) and analysis of variance. The results showed that the background signal intensities of Cy3 channel were higher than those of Cy5 channel. The differences of background signal intensities between the RD-PCR approach and the directly incorporated fluorescently-labeled dNTP labeling were extremely significant (P- Cy3 <0.01, P- Cy5 <0.01). It is concluded that the RD-PCR is a potential and useful sample labeling method with lower background signal intensities for studying high-density long oligonucleotide microarray.