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New Tools for Small RNA Analysis

of miR-124, previously determined by Northern blot analysis using 30 g of total RNA (3). In contrast, only 1 g of total RNA was used in the solution hybridization assay. Moreover, miR-124 expression could be detected in as little as 10 ng of total brain RNA after overnight exposure of the gel (data not shown).

Figure 3. Tissue-specific expression of miR-124 miRNA. miR-124 was detected in 1 g of the indicated FirstChoice Total RNA from 14 different human tissues using the mirVana miRNA Detection Kit and a 5' end-labeled RNA probe prepared with the mirVana Probe & Marker kit. Gel was exposed for 5 hours.


Another application of the mirVana miRNA Detection Kit, is the detection of two or more RNAs in the same sample. For instance, an expressed siRNA and its target mRNA can be monitored simultaneously. See Application: Analysis of miRNA Recovery After Gel Purification (sidebar) or the Ambion miRNA Resource for additional applications of this versatile kit.


Preparation of RNA Probes
Small RNA probes for the solution hybridization assay can be readily prepared using one of two different methods: in vitro transcription (IVT) or 5' end labeling of an existing RNA oligonucleotide. Depending on the probe sequence and the radiolabeled nucleotide used, IVT probes typically result in ~2-fold greater sensitivity than 5' end labeled probes due to their higher specific activity (Figure 4). In contrast, 5' labeled RNA probes are more easily generate
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