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MicroRNA Profiling by Array Analysis Reveals Critical BioMarkers

roducibility

For any method that is used to compare the abundance of RNAs in two different samples, it is critical that the process be highly reproducible. We tested the reproducibility of the miRNA array procedure by repeatedly comparing the miRNA profiles of human prostate and colon samples. Total RNA from a single human prostate sample and a single human colon sample were isolated. The total RNA from the prostate sample was split into six samples, and miRNA from each sample was purified and fluorescently labeled with Cy3 using the mirVana miRNA Labeling Kit. The miRNA from the human colon sample was purified and fluorescently labeled with Cy5.

Each prostate miRNA sample was mixed with one-sixth of the Cy5-labeled colon sample. The six miRNA mixtures were analyzed using a miRNA array containing a complete set of all known human and mouse miRNAs. The signal from the hybridized miRNAs was quantitated, and the signals from the colon and prostate samples were compared. Signal ratios at each element are expressed as a log ratio in Figure 3. The average correlation between the six independent reactions was 98%, indicating that the miRNA array process is highly reproducible.

Figure 3. The miRNA Array Procedure is Reproducible. The reproducibility of the miRNA array procedure was tested by repeatedly comparing the miRNA profiles of human prostate and colon samples. Signal ratios (colon:prostate) at each element are expressed as a log ratio. Standard deviation of Log Ratio between the 6 replicates was determi
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