Fig 1. Reproducibility of Technical Replicates. Single Human Ovary and Heart RNA samples were aliquoted in duplicate to microfuge tubes then processed in parallel through Flash-tag labeling and hybridization to miRBASE version 17 ORB microarrays. Log2-transformed human probe intensities resulting from the replicate hybridizations are plotted.
|miRNA||miRNA Probe Sequence|
Table 1. Example of specificity control probe sequences. Specificity control probes includes a perfect match, single mismatch (MUT1), double mismatch (MUT2), and shuffled (SHUF) version of the probe.
Fig 2. Hybridization Specificity. Signal for specificity control probes complementary to three different microRNAs is shown. ORB quality control specifications require that the signal on a perfect match probe must be at least 100-fold higher than a double-mismatch (MUT2) probe.
Fig 3. Detection of Spiked Ncode microRNAs. 24 attomoles of NCode spike was combined with 2.4 picomoles of LMW RNA prior to labeling for samples 1, 3, 5, 7, 9, 11, 13, 15, 17 but not for samples 2, 4, 6, 8, 10, 12, 14, 16, 18. Probes complementary to the Ncode probe showed greater than 100 fold higher signal in samples where the spike was added, as compared to samples where it was not.
Fig 4. Validation of Microarray data by Real Time qPCR. 22 microRNAs were selected from RT-qPCR validation experiment performed on six different tissue samples (Brain, Kidney, Spleen, Ovary, Heart, and Prostate). The correlation index between Microarray and RT-qPCR was generated and plotted. 21 out of 22 microRNAs showed a high correlation between Microarray and RT-qPCR.