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miRCURY LNA Probe PCR Kits

For high-performance PCR with the miRCURY LNA miRNA Probe PCR System
  • Optimized and dedicated for miRCURY LNA miRNA Probe PCR Assays and Panels
  • Short LNA-enhanced, miRNA-specific PCR components eliminate nonspecific amplification and primer dimers
  • Sensitive and specific performance with the latest QuantiNova enzymes
  • High reaction robustness for room temperature setup and automation
  • Includes the miRCURY Probe PCR Universal Reverse Primer

The miRCURY Probe PCR Kits contain a high-performance PCR master mix specifically designed for probe-based quantification of miRNA using miRCURY LNA miRNA Probe PCR Assays and Panels. Reverse Transcriptase is performed using the miRCURY LNA RT Kit. miRCURY Probe PCR Kits were optimized for use with any real-time cycler.

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Cat No./ID: 339371
miRCURY LNA Probe PCR Kit (200)

For 200 reactions: 2X QuantiNova Probe Master Mix, 10X miRCURY Probe Univ. Primer, Rox Reference Dye, RNase-Free Water

Cat No./ID: 339372
miRCURY LNA Probe PCR Kit (800)

For 800 reactions: 2X QuantiNova Probe Master Mix, 10X miRCURY Probe Univ. Primer, Rox Reference Dye, RNase-Free Water

Cat No./ID: 339373
miRCURY LNA Probe PCR Kit (4000)

For 4000 reactions: 2X QuantiNova Probe Master Mix, 10X miRCURY Probe Univ. Primer, Rox Reference Dye, RNase-Free Water

miRCURY LNA Probe PCR Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.

Product Details

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Reaction stability and setup robustness.
Duplicate reactions using the master mixes of the miRCURY Probe PCR Kit were set up at room temperature. PCRs of one set were run immediately after setup while the second set was stored refrigerated but with light exposure for 24 h. After storage, the PCRs of the second set were initiated. Cq values obtained with the two reaction sets were nearly identical, indicating that PCR performance was not impacted by handling and storage. This allows flexibility in assay workflows.
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Principle of the novel QuantiNova hot-start mechanism.

The QuantiNova DNA Polymerase is kept in an inactive state by the QuantiNova Antibody and QuantiNova Guard until the initial heat activation step.

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miRCURY LNA miRNA Probe PCR System at a glance.
The miRCURY LNA miRNA Probe PCR System uses a single cDNA synthesis reaction for all amplifications, reducing pipetting and saving time and sample. A forward PCR primer and hydrolysis probe that are LNA-enhanced and miRNA-specific deliver highly sensitive miRNA quantification and specificity down to a single-nucleotide sequence difference. The fast and easy workflow takes 2 hours with minimal hands-on time.
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High lot-to-lot consistency.
Strict manufacturing quality standards and robust chemistry ensure consistent performance from one product lot to another, which also translates to high day-to‐day and site‐to‐site reproducibility.
Performance
Unique primer and probe design possibilities

The miRCURY LNA miRNA Probe PCR System is a groundbreaking miRNA detection and quantification system that uses LNA technology to enable new primer and probe designs that greatly enhance assay specificity and sensitivity. On the one hand, the LNA-enhanced primers and probe have higher Tm, which boosts reaction stringency and enables discriminating between miRNAs that differ by a single nucleotide, regardless of AU content. On the other hand, forward primer and probe are miRNA-specific and shorter than in other detection systems (see miRCURY LNA miRNA Probe PCR System at a glance). Thus, assay design is more flexible and smart placement of LNAs greatly improves assay specificity and sensitivity. In fact, the system allows quantifying down to 1 copy of a target miRNA without pre-amplification. Plus, the workflow is fast and simple, going from RNA to data in 2 hours, and exceptional assay robustness allows room temperature setup and delayed runs for greater flexibility and use in automation (see Reaction stability and setup robustness). The QuantiNova enzymes ensure highly reproducible results between technical and biological replicates at the first attempt (see Principle of the novel QuantiNova hot-start mechanism).

Fast and Easy

The easy-to-follow, 2-hour protocol saves time and labor. By using a universal transcription as well as optimized RT-PCR protocols, the procedure is greatly simplified compared to other solutions. Furthermore, the cDNA generated in the RT step can be used across the miRCURY LNA miRNA Probe PCR System, allowing you to seamlessly transition from panels to assays depending on your research needs, and saving sample material and time. The number of pipetting steps is reduced to a minimum, which also minimizes technical variation. This simplified workflow coupled to ISO-compliant assay manufacturing (see High lot-to-lot consistency) makes it possible to achieve extremely high day-to-day and site-to-site reproducibility.

Principle
High-performance reagents for quantitative, real-time RT-PCR

The miRCURY LNA miRNA Probe PCR System enables sensitive, specific miRNA quantification and profiling via real-time PCR. Detection is based on a quencher-labeled hydrolysis probe. The system covers all steps involved in converting miRNA to cDNA and subsequent qPCR detection of miRNAs. The miRCURY LNA miRNA Probe PCR Assays and Panels have been validated using the QuantiNova Probe Master Mix; use of other reagents for qPCR will affect the results.

RNA Spike-In Kit for RT

The RNA Spike-In Kit enables quality control of the RNA isolation, cDNA synthesis and PCR amplification steps of miRCURY LNA miRNA Probe PCR experiments so you can identify any poor-quality samples early in the process and save time and reagents. The kit includes UniSp2, UniSp4 and UniSp5 RNA Spike-in Template Mix and the cel-miR-39-3p RNA Spike-in Template.

miRCURY LNA RT Kit

One universal reverse transcription kit for two detection systems. The miRCURY LNA RT Kit is used for both the miRCURY LNA miRNA PCR System, which employs SYBR® Green for detection, and the probe-based miRCURY LNA miRNA Probe PCR System. The kit includes two system-specific reaction buffers and all the necessary reagents to perform fast and convenient miRNA polyadenylation and reverse transcription in a single reaction step.

miRCURY Probe PCR Kit

This kit contains the high-performance QuantiNova Probe Master Mix and the miRCURY Probe PCR Universal Reverse Primer. Combined with a miRCURY LNA miRNA Probe {CR Assay or a miRCURY LNA miRNA Probe PCR Panel, the chemistry enables miRNA detection via quantitative real-time PCR.

miRCURY LNA miRNA Probe PCR Assays

These primer and probe assays use miRNA-specific hydrolysis probes and miRNA-specific forward primers designed to detect mature miRNAs using the most up-to-date sequence information from miRBase. The miRCURY Probe Universal Reverse Primer is contained in the miRCURY Probe PCR Kit.

miRCURY LNA miRNA Probe PCR Panels

Read-to-use, predesigned, single-use PCR panel plates to rapidly profile mature miRNAs focusing on the miRNome, specific pathways, certain sample types like serum/plasma, or in a user-designed custom panel. 

GeneGlobe Data Analysis Center

This complimentary, intuitive web-based analysis tool guides you step-by-step via an easy wizard through the normalization and analysis of your data and generates publication-ready figures.

Procedure

The first step, the reverse transcription reaction, is an easy-to-perform, single-tube reaction. Total RNA, including miRNA, is used as starting material and the resulting cDNA is used in the real-time PCR amplification with LNA-enhanced probe and primer to detect the mature miRNAs. This enables straightforward miRNA quantification and profiling, accessible to any lab with a real-time PCR cycler.

Applications

The miRCURY Probe PCR Kits are used as part of the miRCURY LNA miRNA Probe PCR System for:

  • miRNA expression detection, profiling and quantification
  • miRNA biomarker development – screening and identification
  • miRNA regulation and pathway analysis
  • miRNA detection in complex sample types and samples with low miRNA expression levels
  • snoRNA detection
  • Validation of miRNA sequencing

Product Resources

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Kit Handbooks (2)
For highly sensitive, ultrafast real-time RT-PCR detection of miRNAs from exosomes, serum/plasma, and other biofluids
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For highly sensitive, real-time RT-PCR detection of miRNAs using hydrolysis probes
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