microRNA Profiling by qPCR
microRNA qPCR Arrays
High-throughput microRNA Expression Profiling
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.
miProfile miRNA qPCR Arrays
The miProfile miRNA PCR Arrays are designed for profiling the expression of pre-defined sets of miRNAs, such as whole genome, cancer-related, disease-related, stem cell-related or exosome-derived miRNAs, in various tissues or cells. The cancer-related or disease-related miRNAs are carefully chosen for their close correlation with a specific cancer or disease based on a thorough literature search of peer-reviewed publications. The resulting differential expression 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.
Universal real-time PCR conditions were developed for easy profiling and analysis of 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.
- Profile whole genome, cancer-related or disease-related microRNAs
- Validated primers designed with proprietary algorithm
- Detect microRNA from as little as 10 pg of small RNA or 20 pg of total RNA
- Distinguish miRNAs with single nucleotide mismatches. Each primer set has been experimental validated for specific amplification
- Simultaneously detect microRNAs at different expression levels
- High reproducibility (R2 > 0.99) for inter-array and intra-array replicates
96-well Array Format
Five 96-well qPCR array formats (A, B, C, D, and E) are suitable for use with the following real-time PCR instruments are provided. Before placing an order, please take care to choose the right kit version which is compatible with your instrument.
384-well Array Format
Three 384-well qPCR array formats (F, G, and H) are suitable for use with the following real-time PCR instruments are provided. Before placing an order, please take care to choose the right kit version which is compatible with your instrument.
How It Works
microRNA qPCR Array Experimental Workflow
microRNA RT-PCR Mechanism
Performance Data
Linear Range and Sensitivity (Total RNA)
Broad linear range and high sensitivity. Starting with serially diluted amounts of human colon cancer total RNA, miR-21 and miR-1260 were detected using All-in-One™ miRNA qRT-PCR Detection Kit. The resulting Ct values were plotted against the log10 of the amounts of input total RNA. The data demonstrated a broad linear dynamic range from 20 pg to 2 µg of input total RNA as well as high sensitivity. This allows the detection of miRNAs at varying expression levels, including low expressers.
Inter-Array Reproducibility
High inter-array reproducibility. Two miProfile PCR array replicates (plate A and B) were analyzed using human total RNA (10-tissue mix) on the Bio-Rad iQ5. The Ct values of the replicate plates were plotted against each other. R2 > 0.99 were observed for high inter-array reproducibility.
Single Nucleotide Mismatches
Specificity of miRNA detection. miRNA miR-29a and miR-29c with one single nucleotide mismatch (B) can be distinguished. Relative detection, defined as a percentage of the perfect match (100% x 2 - ΔCt), was calculated using the Ct values of on-target and off-target assays, which were performed to detect miRNA plasmid DNA templates using All-in-One miRNA qRT-PCR Detection Kits (A).