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Highly Sensitive microRNA Array Performance


We also tested the reproducibility of our miRNA array system by comparing miRNA profiles obtained from human colon vs. human prostate in six independent array experiments. Total RNA was isolated from a single human prostate and colon sample. Each total RNA sample was split into six replicates. miRNAs in each sample were independently isolated and labeled. Each of the six Cy3-labeled prostate miRNA samples was combined with a Cy5-labeled colon sample and the six prostate/colon pairs were hybridized to miRNA arrays. The Log2(R/G Normalized Ratio) of each spot is shown in Figure 3B. The average correlation between the six independent experiments was 98%, indicating that the miRNA array process was highly reproducible.


Accurate Data

The most important measurement for a microarray procedure is whether the quantitative array data truly reflect the abundance of miRNAs in total RNA samples. We set up an extensive experimental analysis using several tissue samples to address accuracy. miRNA was purified from FirstChoice Total RNA from human bladder, lung, and uterus.

The miRNAs were labeled and hybridized with arrays to compare the expression of more than 160 miRNAs in bladder, lung, and uterus.

The relative abundance of seven different miRNAs was measured by Northern analysis or a hybridization ass
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