Systematic comparison of the fidelity of aRNA, mRNA and T-RNA on gene expression profiling using cDNA microarray.

摘要: In cDNA microarray technology, there are three main reverse transcription based RNA labeling methods, using total (T-RNA), mRNA, and amplified antisense (aRNA), respectively. However, despite the common use of types RNAs, limited data available regarding their differences concordances. this report, we compared methods through two sets self-comparison experiments same sample in all cases. Within each method, duplicate hybridizations highly reproducible with low biases, which randomly produced. When combining different RNAs within a single array, correlation coefficients between channels rather low, while discrepancies persistent. Furthermore, fidelity aRNA mRNA microarrays expression profile study shows no significant difference standard T-RNA methods. These results suggest that some abundance selectively changed during amplification/mRNA purification processes, but it will not affect gene ratio samples if type used. Therefore can be used profiling analysis as long test reference generated by identical method study.