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New Tools for Small RNA Analysis

probe should carry only a few nontarget-specific nucleotides at its 3' end. The kit can also be used to label and purify DNA probes (see Northern Blot in Figure 4) and to prepare small Decade RNA markers (150, 100, 90, 80, 70, 60, 50, 40, 30, 20, and 10 nt; see Figure 5), which are included in the kit.

Figure 5. miRNA Detection after Gel Purification. miRNAs were gel purified on a 15% denaturing polyacrylamide gel from 150 g of mouse lung or thymus total RNA using standard procedures. Recovery was analyzed with the mirVana miRNA Detection Kit and autoradiography (2 hours exposure) by comparing the amount of miR-16 miRNA in the sample before (1% or 0.5% of the starting total RNA) and after purification (2% or 1% of the miRNA fraction). Percentage of recovery (59 and 53% respectively) was then determined by quantification with a phosphorimager. mir-16 probe and Decade Markers were prepared with the reagents included in the mirVana Probe & Marker Kit.


A Complete Solution
Ambion's mirVana system provides the first complete solution for small RNA analysis. The system allows for the isolation of total RNA samples that contain small RNAs--or the preparation of RNA samples that are enriched for small RNAs. Alternatively, "miRNA certified" total RNAs from over 80 different human, mouse, and rat tissues can simply be purchased. Once the RNA samples are obtained, high specific activity radiolabeled probes can be prepared by IVT or by 5' end-labeling, and target small RNAs be detected using
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