For exceptionally sensitive and specific miRNA profiling using LNA-enhanced, SYBR® Green-based miRNA qPCR

Features

Unmatched sensitivity enables profiling 752 miRNAs using only 40 ng total RNA

Available for human, mouse and rat miRNAs

Fast and easy to use with no pre-amplification required

Reliable discrimination of closely related miRNAs and mature and precursor miRNAs

Product Details

miRCURY LNA miRNA miRNome PCR Panels enable exceptionally sensitive and specific miRNA expression profiling using LNA technology and the miRCURY LNA miRNA PCR System. These predesigned PCR primer sets are pre-aliquoted in 384-well PCR plates and are ready for use. Available for human, mouse and rat miRNAs, the miRCURY LNA miRNA miRNome PCR Panels allow you to profile up to 752 human or rodent miRNAs using only 40 ng total RNA, with no required pre-amplification.

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Performance

Top performance in specificity

The miRCURY LNA miRNA PCR platform is the only miRNA quantification platform with absolute specificity and out-performed another probe-based miRNA qPCR platform in specificity tests . This eliminates false positives and ensures only robust and reliable miRNA signals. 

Fast and reproducible results from minimal sample amounts

The miRCURY LNA miRNA miRNome PCR Panels enable profiling of 752 miRNAs from just 40 ng of total RNA, so you can perform high-quality miRNA expression profiling in samples that contain very little total RNA, such as FFPE sections and serum/plasma .

The easy-to-follow, 3-hour protocol saves you both time and effort in the laboratory. Simply dilute the synthesized cDNA, combine it with the miRCURY LNA SYBR Green Master Mix and aliquot into the 384-well PCR plate, which is then ready to run in your real-time PCR instrument. The number of pipetting steps is reduced to a minimum, reducing technical variation. As a result, you can achieve extremely high reproducibility from day-to-day and even site-to-site. High reproducibility is even possible when profiling difficult samples, such as RNA purified from serum .

Principle

Features

The miRCURY LNA miRNA miRNome PCR Panels enable exceptionally sensitive, high-throughput expression profiling from minimal amounts of starting material. The combination of a universal cDNA synthesis reaction and ready-to-use PCR panels provides fast and easy miRNA profiling. The panels are provided in 384-well plates that contain dried-down primers for one reaction per well and simply require addition of cDNA and miRCURY LNA SYBR Green Master Mix prior to real-time PCR amplification.

The LNA-enhanced, miRNA-specific PCR primers offer exceptional sensitivity and extremely low background, enabling accurate quantitation of very low miRNA levels. Plus, high specificity allows discrimination between closely related miRNA sequences, with a quick and easy protocol that gives you results in just 3 hours. 

Coverage

Two panels covering 752 human miRNAs and two panels covering 752 mouse and rat miRNAs are available. Panel I for human and mouse/rat contains high-priority miRNA primer sets. These miRNAs are generally more highly expressed, more likely to be differentially expressed in disease or more often cited in the literature. Panel II contains primer sets for the most important miRNAs not available on Panel I. To view the list of primer sets in each panel, download the plate layout files.

The miRNome PCR Panels also include 6 potential reference gene primer sets – three small RNA reference genes (U6, SNORD38B and SNORD49A for human and U6 snRNA, RNU5G, RNU1A for mouse and rat) and three miRNA reference genes (miR-103-3p, miR-191-5p and hsa-miR-423-5p or mmu-miR-423-3p) – all found on Panel I. Panel I also contains qPCR assays for the 5 synthetic RNAs in the RNA Spike-in Kit (cel-miR-39-3p, UniSp2, UniSp4, UniSp5 and UniSp6) that may be used for monitoring the cDNA synthesis or RNA purification. Finally, each panel contains an inter-plate calibrator in triplicate and an empty negative control. 

Version update

The updated miRCURY LNA miRNA miRNome PCR Panels include 54 updated assays on Panel I and 33 updated assays on Panel II. The assays target the same miRNAs as v3 panels. In addition, the two spike-in primer sets UniSp5 and UniSp6 have been updated for improved performance. Their respective target templates will be the same.

Procedure

The easy-to-follow, 3-hour protocol saves you both time and effort in the laboratory. Simply dilute the synthesized cDNA, combine it with the miRCURY LNA SYBR Green master mix and aliquot into the 384-well PCR plate containing the pre-aliquoted miRCURY LNA miRNA miRNome Panels. The plate is now ready to run in your real-time PCR instrument.

Applications

miRNA quantification in serum, plasma, exosomes, urine and other liquid biopsies; biomarker discovery