Prepare siRNA and miRNA Probes in Just 1 Hour
- Prepare your probe in less than 1 hour
- Ideal for siRNA/miRNA studies
- Synthesize high specific activity RNA probes for RPA, Northern, Southern, or in situ hybridization
- No cloning required
With the mirVana miRNA Probe Construction Kit, short (<100 nt) radiolabeled or nonisotopically labeled RNA probes can be generated in less than 1 hour. These probes are ideal for the investigation of a variety of small RNAs. Radiolabeled probes prepared with the mirVana miRNA Probe Construction Kit have been successfully used for the detection of small nuclear RNA (snRNA), small interfering RNA (siRNA), micro RNA (miRNA), and mRNA by RPAs, Northerns and in situ hybridization. Nonisotopically labeled probes generated with the kit have also been used to study the distribution of miRNA or mRNA in tissues by in situ hybridization (See Analysis of miRNA Distribution in Mouse Brain, right).
RNA Probe Synthesis Made Fast, Simple and Inexpensive
Complete Kit for Probe Preparation
Figure 1. miR-16 and miR-22 Expression in Mouse Brain. In situ hybridization was performed on coronal mouse forebrain sections. In situ hybridized cells in the mouse brain cortex (A,B,D,E; 20X magnification) and the head of the caudate nucleus (C,F; 40X magnification) are indicated by arrows. Nonisotopically labeled probes were prepared with Ambion's MAXIscript™ Kit (1.5 kb probe) or with the mirVana miRNA Probe Construction Kit (32 nt long probes, 22 nt target specific sequence).
Ambion's scientists have optimized procedures for the sensitive and specific detection of mRNA and miRNA by in situ hybridization with small probes. As shown in Figure 1 (below left), the same cell-specific expression of VIP mRNA was observed in mouse cortex with a 1.5 kb probe as with a 22 nt long probe prepared with the mirVana miRNA Probe Construction Kit (Panels A and B). This result demonstrated that the small probes yielded specific signal.
Using additional small RNA probes, miR-16 and miR-22 miRNAs were found to be widely distributed in the cortex layers 2 and 3 (Panels B and E). Consistent with this result, high levels of miR-16 and miR-22 expression were also detected in mouse brain total RNA by hybridization in solution with the mirVana miRNA Detection Kit (data not shown). However, differences between the miR-16 and miR-22 expression patterns were noted within the brain. For example, only very few miR-16 positive cells were observed in the head of the caudate nucleus (Panels C and F). The staining pattern in this area was characteristic of cytoplasmic subcellular localization for both miRNA targets.