• Rapid procedure — prepare transcription templates and synthesize RNA probes in less than 1 hour
  
  
• Ideal for siRNA/miRNA studies
  
  
• Compatible with the mirVana™ miRNA Detection Kit, Northern analysis, and in situ hybridization
  
  
• No cloning required
  
  
• More economical than labeling chemically synthesized RNA
  
  
• Produces probes with higher specific activity than 5' end labeling, resulting in greater sensitivity
  
  

For a faster and more sensitive alternative to Northern analysis, please see our mirVana™ miRNA Detection Kit.

With the mirVana miRNA Probe Construction Kit, short (<100 nt) radiolabeled or nonisotopically labeled RNA probes can be generated by in vitro transcription in less than 1 hour. 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. In addition, because the RNA oligonucleotide is synthesized enzymatically, small RNA probes prepared by IVT are less expensive than probes prepared by chemical synthesis followed by 5' end labeling.

Radiolabeled probes prepared with the mirVana miRNA Probe Construction Kit have been successfully used for the detection of microRNA (miRNA), small nuclear RNA (snRNA), and small interfering RNA (siRNA), as well as for mRNA. 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.

RNA Probe Synthesis Made Fast, Simple and Inexpensive
The mirVana miRNA Probe Construction Kit includes reagents for preparation of in vitro transcription templates as well as reagents for in vitro transcription. For template preparation, all you need to supply is a short, inexpensive DNA oligonucleotide specific for the target of interest that also includes an 8 base sequence complementary to the 3' end of the T7 promoter primer included in the kit. This oligonucleotide is annealed to the provided T7 promoter primer, and a 30 minute fill-in reaction with Klenow fragment generates the double-stranded transcription template. The resulting template is simply mixed with the provided T7 RNA Polymerase, rNTPs, and Transcription Buffer along with your choice of labeled rNTP. The reaction is then incubated for as little as 10 minutes at 37°C to generate labeled probe.

Complete Kit for Probe Preparation
The kit includes enough reagents to prepare 30 dsDNA templates, and to perform 30 in vitro transcription reactions. Each template construction reaction supplies sufficient template for up to 20 transcription reactions. The kit also includes a functional positive control, DNase I to remove residual DNA template, and Probe Elution Buffer to gel purify RNA transcripts.

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