( with the highest expression levels evid...)Detecting miRNA & siRNA

with the highest expression levels evident in lung and thymus. These variations across tissues were confirmed by Northern blot analysis with the same mir-16 probe. Interestingly the mir-22 probe showed a completely different pattern of expression. miR-22 was highly expressed in lung and ovary and was present in spleen, thymus, and testicle at levels that would not have been detectable with standard Northern blotting techniques (data not shown). The relative abundance of miR-16 and miR-22 miRNA in mouse lung was also confirmed by multi-target detection with the simultaneous use of the mir-16+4 and mir-22 probes in the assay (Figure 3).

Figure 4. miR-16 and miR-22 Expression in Mouse Tissues. miR-16 and miR-22 miRNAs (22 nt) were detected in 1 g of FirstChoice Total RNA from five different mouse tissues using 32 nt long mir-16 or mir-22 probes generated with the mirVana miRNA Probe Construction Kit. The same differential expression of miR-16 across tissues was observed by Northern blot analysis (2 days exposure) or by hybridization in solution (2 hr exposure). RNAs were analyzed on 15% denaturing polyacrylamide gels. As a loading control, the same RNA samples were resolved on a 1.2% denaturing agarose gel and U1 snRNA expression was analyzed by Northern blot.

Detecting Small RNAs and mRNAs in the Same Sample
In many cases, it may be desirable to simultaneously detect small RNAs such as miRNA or siRNA with one or more mRNAs. To determine if the assay was compatible with this application, an antisense probe to GAPDH mRNA was designed to generate a protected RNA fragment slightly lo
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