For high-performance PCR with the miRCURY LNA miRNA Probe PCR System

Features

•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

Product Details

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.

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.

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