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miRCURY LNA miRNA miRNome PCR Panels

For exceptionally sensitive and specific miRNA profiling using LNA-enhanced miRNA qPCR
  • 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

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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Ready-to-Use miRNome PCR panels of miRNA assays in 384-well format (Human, Mouse&Rat)
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339322 YAHS-301Y varies
Ready-to-Use miRNome PCR panels of miRNA assays in 384-well format (Human, Mouse&Rat)
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339322 YAHS-312Y varies
Ready-to-Use miRNome PCR panels of miRNA assays in 384-well format (Human, Mouse&Rat)
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339322 YAMR-301Y varies
Ready-to-Use miRNome PCR panels of miRNA assays in 384-well format (Human, Mouse&Rat)
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339322 YAMR-312Y varies
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miRCURY LNA miRNA miRNome PCR Panels are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.


0
Expression profiling of 730 miRNAs using 40 ng total RNA from tumor and normal FFPE sections.
Real-time PCR was performed using triplicate RT reactions per sample on miRCURY LNA miRNome Human Panels I and II (730 miRNAs in total). Data from 424 miRNAs with Cp values less than 37 are included. Normalized expression is shown as fold changes in tumor compared to normal. Out of 424 miRNAs expressed, 144 were more than 2-fold down-regulated in the tumor (blue dots) and 26 were more than 2-fold up-regulated in the tumor (red dots).
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Expression profiling of 368 miRNAs using total RNA from 35 μl serum.
Real-time PCR was performed using triplicate RT reactions on total RNA purified from serum. Average Cq (quantification cycle) values from the miRCURY LNA miRNome Human PCR panel I are shown (368 miRNAs). Over 120 miRNAs showed robust expression with Cq values below 35. hsa-miR-16, which is known to be highly expressed in serum/plasma, and hsa-let-7a are indicated in red. The U6 reference gene is indicated in orange.
2
Specificity test alongside competitor probe-based miRNA qPCR system.
Specificity was assessed using 8 pools each containing at least 80 different synthetic miRNAs. Closely related miRNAs are placed in different pools. This example shows results from the hsa-miR-1 assay. The miRCURY LNA miRNA PCR System performs perfectly with a high signal in pool 1, which contains the hsa-miR-1 synthetic miRNA target, and only a borderline signal in pool 7, which contains the most closely related miRNA (hsa-miR-206). The competitor platform uses ligation-based cDNA synthesis and only one miRNA-specific probe. Competitor T shows a lack of specificity. The highest signal for Competitor T is obtained with pool 1. However, non-specific false-positive signals are obtained from all other pools in the absence of hsa-miR-1 template, within a Cq range that would be interpreted as real signals.
3
Excellent site-to-site correlation.
Aliquots of the same heart and liver total RNA were expression profiled using miRCURY LNA miRNA PCR Panels in two different locations (on separate days by different operators) using different Roche LightCycler 480 Real-Time PCR instruments. The correlation between average Cq values (triplicate RT reactions) from all miRNAs with signals below 35 Cq values (total of 297 datapoints) is shown.
4
Excellent reproducibility between RT reactions on total RNA from serum.
Raw Cq values from two separate RT reactions (RT1 and RT2) on total RNA purified from 65 µl serum are shown. A total of 730 miRNAs were profiled. Only miRNAs with Cq values below 35 have been included (133 data points).
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Excellent day-to-day reproducibility.
Different RT reactions using 40 ng heart and liver total RNA were profiled on the miRCURY LNA miRNome Human PCR Panel I and II on different days. The correlation between raw Cq values from all miRNAs with signals below 35 Cq values is shown (total of 297 data points).
Performance
Top performance in specificity
The miRCURY LNA miRNA PCR platform is the only miRNA quantification platform with absolute specificity (see the miRQC study published in Nature Methods) and out-performed another probe-based miRNA qPCR platform in specificity tests (see figure Specificity test alongside competitor probe-based miRNA qPCR system). 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 (see figures Expression profiling of 730 miRNAs using 40 ng total RNA from tumor and normal FFPE sections and Expression profiling of 368 miRNAs using total RNA from 35 μl serum).

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 (see figures Excellent day-to-day reproducibility and Excellent site-to-site correlation). High reproducibility is even possible when profiling difficult samples, such as RNA purified from serum (see figure Excellent reproducibility between RT reactions on total RNA 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
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