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miProfile™ miRNA qPCR Arrays

Introduction

For high-throughput profiling of miRNA expression

 

MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression at the post-transcriptional level. Usually 21-23 nucleotides in length, miRNAs are important modulators in cellular pathways and are highly conserved in eukaryotic organisms. Irregularities in miRNA-regulated gene expression have been found to be associated with cancers, cardiovascular disorders and a variety of other diseases.

The miProfile miRNA PCR Arrays are designed for profiling the expressions of pre-defined or customized sets of miRNAs in various tissues or cells. The resulting differential expressions of profiled miRNAs help researchers to identify those miRNAs that are of biological significance and importance relevant to their research. Each 96-well plate contains up to 84 pairs of PCR primers (forward: miRNA-specific primer; reverse: universal primer) and 12 wells of different types of controls, which are used to monitor the efficiency of the entire experimental process – from reverse transcription to qPCR reaction.

Each miRNA-specific primer used in the qPCR arrays has been experimentally validated to yield a single dissociation curve peak and to generate a single amplification of the correct size for the targeted miRNA. A cDNA pool, containing reverse transcript products of 10 different tissue total RNAs, was used as the qPCR validation template.

Using a universal real-time PCR condition, one can easily profile and analyze the miRNA expression in a high-throughput fashion.The All-in-One™ miRNA First-Strand cDNA Synthesis Kits and qPCR Mix Kits are the recommended  and supported RT-PCR reagents for use with the miProfile miRNA qPCR arrays. These reagents have been optimized to produce high sensitivity, efficiency, and specificity.

Key advantages

Validated miRNA primers

  • Each miRNA primer is designed using a proprietary algorithm and experimentally validated

Robust performance

  • Sensitive – Detect miRNA from as little as 10 pg of small RNA or 20 pg of total RNA
  • Specific – Be able to distinguish miRNAs with single nucleotide mismatches. Each primer set has been experimental validated for specific amplification
  • Broad linearity– Allow miRNA at varieties of expression level to be detected simultaneously
  • Reproducible – High reproducibility (R2>0.99) for inter-array and intra-array replicates

Genome-wide coverage, pre-arranged groups, or customized groups

  • Largest genome-wide miRNA coverage
  • Cancer-related groups
  • Customized miRNA arrays for focused study

 

Figure 1. miRNA PCR array experimental work flow (left) and miRNA RT-PCR mechanism (right).

 

Performance data

 

Linear Range and Sensitivity (Total RNA)

   
 

Figure 2. Broad linear range and high sensitivity

     

Inter-Array Reproducibility

   
 

Figure 3. High inter-array reproducibility 

     

Single Nucleotide Mismatches

   
 

Figure 4. Specificity of miRNA detection