PCR

miRNA quantification

Take a deep and exact look at any miRNA

miRNA quantification is a process used to measure the expression levels of microRNAs (miRNAs), which are small non-coding RNAs involved in gene regulation. It provides insights into various biological processes and diseases.

Quantitative PCR (qPCR) is the tool of choice for analyzing known miRNA targets, offering several benefits over NGS. qPCR is fast, simple and cost efficient, providing extremely sensitive analyses from low amounts of input RNA. It also easily scales from screening and profiling to validation of NGS studies and verification of results from functional studies.

However, miRNA qPCR presents special challenges due to their low abundance, short sequences, large variation in GC content and highly homologous families, making the design of very specific primers and probes difficult. Our miRCURY LNA miRNA PCR and Probe PCR systems are designed to overcome these issues, ensuring robust and highly sensitive miRNA profiles, even for your most demanding applications. Trust our solutions for precise miRNA quantification, and experience the reliability and sensitivity you need for your research.

LNA-enhanced miRNA qPCR

How you can benefit

Analyze any miRNA biomarker with ease

miRNAs show high potential as biomarkers for diseases and biological processes, such as cancer, heart disease, nervous system function, hemostasis, DNA repair and others.

Our offering of predesigned miRNA PCR assays is the most comprehensive on the market, so you can easily find the assays you need for your targets of interest. Each of our portfolios includes over 31,000 unique assays for 271 different species and provides full coverage of targets in miRBase 22.

Are you studying a novel miRNA or small RNA? Use our convenient online builder to order custom assays if no predesigned assay is available.

See more in GeneGlobe

Study the relevant targets for relevant results

Our predesigned focus panels cover multiple species and provide a fast and convenient way for screening biologically relevant miRNAs. The targets on each panel were carefully selected from a mix of literature searches, NGS and other studies and advice of leading experts.

The serum/plasma focus panels were meticulously developed based on our extensive analyses of miRNA expression in blood, serum and plasma samples, as well as on the limited number of peer-reviewed published papers available. Over 1 million in-house and collaborative data points from serum/plasma samples from healthy and diseased individuals were used in selecting relevant miRNAs – including miRNA expression data from different stages of various cancers, neurological disorders, neurodegenerative diseases, allergies, inflammation, etc.

All panels can be easily ordered online through GeneGlobe.

Explore focus panels

Optimally designed assays for any miRNA target

Short sequence lengths and high similarities complicate miRNA assay design. We overcome the difficulty by optimizing our assays with locked nucleic acids (LNA).

LNA bases increase primer binding affinity, allowing us to design very short primers and probes, which can be placed directly on the miRNA target to create two miRNA-specific annealing points that do not overlap. This just isn't possible with standard oligos. The result is an optimal assay design that accurately detects your miRNA of interest with a specificity that can even distinguish single nucleotide differences.

Plus, the LNA makes it possible to normalize the primer Tm so that all primers have the optimal Tm for uniform amplification. This lets you simultaneously detect both high- and low-abundance targets with heterogeneous GC content.

Quantify low-abundance miRNAs without preamplification

Quantification of low-abundance miRNAs by qPCR usually requires preamplification, but the miRCURY LNA miRNA PCR systems provide accurate results without this extra step.

What’s the secret? It’s the LNA enhancement. The increased binding affinity of LNA oligos allows us to design assays that can detect any biologically relevant miRNA with the highest specificity and sensitivity – without the need for preamplification.

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Workflow & products

Your choice of detection chemistry

We offer you the choice between SYBR-Green and hydrolysis probe-based miRNA detection, both providing highly sensitive and specific detection.

Which detection chemistry is right for you?

Each method offers unique advantages when it comes to cost, ease of use, specificity, robustness, design flexibility and multiplexing capacity.

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Fast and simple workflow

The miRCURY LNA miRNA PCR systems combine the speed and convenience of a universal reverse transcription (RT) reaction with the sensitivity and specificity of LNA-enhanced qPCR primers. The simple workflow delivers fast and accurate results for all sample types, including tissues, cells, exosomes and liquid biopsies.

What you need to get started

A universal reverse transcription kit (1) contains the chemistry for both SYBR Green and probe-based detection. Dedicated PCR mastermixes (3) for each system are combined with your choice of pre-designed or self-designed assays or panels (4). The optional RNA Spike-In Kit (2) lets you perform quality control of the RNA isolation, cDNA synthesis and PCR amplification steps, so you can identify any poor-quality samples early on, eliminate them from your study and save time and reagents.

	Detection system	SYBR Green	LNA-enhanced hydrolysis probe
1	Reverse Transcription	miRCURY LNA RT Kit	miRCURY LNA RT Kit
2	Optional spike-in	RNA Spike-In Kit, For RT	RNA Spike-In Kit, For RT
3	Mastermix for qPCR	miRCURY LNA SYBR Green PCR Kits	miRCURY LNA Probe PCR Kits
4	Assay or panel	• miRCURY LNA miRNA PCR Assays • miRCURY LNA miRNA Custom PCR Assays • miRCURY LNA miRNA Focus PCR Panels • miRCURY LNA miRNA Custom PCR Panels • miRCURY LNA miRNA miRNome PCR Panels	• miRCURY LNA miRNA Probe PCR Assays • miRCURY LNA miRNA Custom Probe PCR Assays Coming soon: • miRCURY LNA miRNA Focus Probe PCR Panels • miRCURY LNA miRNA Custom Probe PCR Panels • miRCURY LNA miRNA miRNome Probe PCR Panels

Which assay or panel is best for your study?

The miRCURY LNA miRNA PCR systems let you tailor the experimental setup to various types of studies with individual assays or fully flexible custom panels. You can use the same cDNA reaction and shift seamlessly between individual primers, predesigned miRNome and Focus panels or custom panels.

Product Type	Advantage	Type of Study
Starter kit*	Few assays with a low number of samples	Getting started
Individual assays	Economical option for small-scale screening of a few miRNAs or for quantification of a high number of samples	Validation experiment with few miRNAs (pilot/preliminary experiment)
Custom panels	Pre-pipetted, custom-selected assays ensure high reproducibility	Focused screening or validation experiment
Focus panels	Pre-defined assay selections targeting certain research areas	Focused screening
miRNome panels	Cover all important miRNAs	Genome-wide screening

Products

Resources

Dive deeper into webinars and other resources

Get great results, even from difficult sample types

Circulating miRNAs in liquid biopsy samples such as serum/plasma, urine and exosomes have broad potential as disease biomarkers. However, these samples contain low amounts of RNA, high levels of inhibitors and are susceptible to many preanalytical variables, making miRNA profiling a challenge.

Our biofluid miRNA guidelines provide expert information and tips for biomarker discovery and validation in biofluid samples. Learn about optimized sample handling and preparation, quality control methods and miRNA detection techniques to ensure biofluid miRNA profiling success.

Get the guidelines

miRCURY LNA performs best in miRQC study

In the largest miRNA benchmarking study to date, Pieter Mestdagh and colleagues compared the performance of commercially available miRNA profiling platforms in key areas. The miRCURY LNA platform was the only top performer in all areas, combining both high sensitivity and specificity. Access the article here.