• A synthetic equimolar pool of most known miRNAs in human, mouse, rat genomes
  
  
• Useful as positive control and quantitative standard for real-time PCR, array hybridization, and sample preparation
  
  
• Based on miRBase v9.1 hosted by the Sanger Institute
  
  

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A Positive Control and Quantitative Standard for miRNA Analyses
The mirVana™ miRNA Reference Panel v9.1 is an equimolar pool of synthetic RNA oligonucleotides representing most of the human, mouse, and rat miRNAs in miRBase Sequence Database Version 9.1. This miRNA Reference Panel is intended for use as a positive control and/or quantitative standard in a variety of miRNA-related applications.

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Suitable for a Variety of Applications
The mirVana miRNA Reference Panel is suitable for use as a positive control and/or quantitative standard in such miRNA applications as array analysis, real-time PCR, sample preparation, and small RNA cloning/sequence analysis. In addition to use as a hybridization contol for Ambion mirVana miRNA Bioarrays, the miRNA Reference Panel can be used in conjunction with the mirVana miRNA Probe Set on spotted arrays to control for printing variation. In both cases this allows for tighter control of array processing and data normalization methods. The mirVana miRNA Reference Panel is also useful as a quantitative standard for real-time PCR due to the careful mixing of equimolar amounts of each oligonucleotide in the panel. This allows for absolute quantification of molar or mass amounts of miRNAs in an unknown sample relative to a standard curve generated using this product (Figure 1). In addition, Applied Biosystems scientists have demonstrated the usefulness of the miRNA Reference Panel as an in-process control for sample preparation using the flashPAGE™ Fractionator Apparatus and other mirVana sample preparation methods (e.g., the mirVana miRNA Isolation Kit; data not shown). The mirVana miRNA Reference Panel can also serve as a positive control for small RNA cloning and sequence analysis. Researchers conducting such experiments may wish to assay a radiolabeled probe for a target miRNA within miRBase v9.1 against this panel to serve as a positive control in Northern blotting.

Linearity of Dose Response
Use of the mirVana miRNA Reference Panel as a real-time PCR and array hybridization control is demonstrated in Figures 2 and 3. The experiment shown was designed to test the linearity of dose response as a function of increasing mass for the two methods of analysis. The mass inputs of synthetic miRNAs spanned the range of detection for mirVana™ miRNA Bioarrays from background to saturating levels. Both methods showed a strong linear response for most sequences across the subpools used, and the concordance between the two methods was high (Figure 2). For real-time PCR the highest mass inputs yielded CT values of about 10, and the lowest non-zero inputs yielded CT values of approximately 23, providing an even greater dynamic range and sensitivity of miRNA detection (Figure 3). This experiment shows that the synthetic miRNAs of the mirVana miRNA Reference Panel behave in a predictable manner and would, therefore, be useful as a positive control and quantitative standard for analysis of unknown miRNA samples.

The mirVana miRNA Reference Panel is comprised of unmodified, HPLC-purified, single-stranded RNA oligonucleotides. One miRNA Reference Panel is sufficient to serve as a positive control for 100 TaqMan® Real-Time Assays or 20 mirVana miRNA Bioarrays.

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