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miRCURY LNA miRNA Custom PCR Panels

Design your own custom-formatted 96- and 384-well qPCR plates for the miRCURY LNA miRNA PCR System
  • Combine predesigned verified and custom-designed LNA PCR Assays in single-use, ready-to-use plates
  • Customize for your favored plate type, layout, PCR instrument and batch size
  • Great for sensitive, specific and reproducible miRNA profiling of a medium to large number of samples
  • Use our new easy-to-use online design tool

miRCURY LNA miRNA Custom Panels are ready-to-use 96- and 384-well plates containing individual miRCURY LNA miRNA PCR Assays according to your particular requirements. Both forward and reverse PCR amplification primers are miRNA-specific and are optimized with LNA technology, enabling extremely sensitive and specific miRNA quantification with the miRCURY LNA miRNA PCR System. Each well contains sufficient primer set for one 10 µl reaction.

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Product Product no. Cat. no. List price:
miRCURY LNA Custom PCR Panel (1)
miRCURY LNA Custom PCR Panel provided in one ready-to-use 96- or 384-well plate; each well contains primers sufficient for one 10 µl reaction
339330 Varies Create and order
miRCURY LNA Custom PCR Panel Additional Plates (4)
Additional miRCURY LNA Custom PCR Panel plates; only available in addition to the base plates ordered through the core product (cat. no. 339330)
339332 Varies Create and order

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


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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.
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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.
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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 System 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.

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 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
Design your own 96- or 384-well miRNA qPCR plates based on a fully flexible layout and several convenient, predefined layouts. Place predesigned miRCURY LNA miRNA PCR Assays or your own Custom miRCURY PCR Assays into the panel using an intuitive and easy-to-use online plate configurator. The panels are delivered ready-to-use, with one single assay per well for a 10 µl reaction volume. Just add cDNA and miRCURY LNA SYBR Green Master Mix.

Coverage
Custom miRNA PCR Panels are available in 2 different formats:
  • 96-well, ready-to-use: design your panels from one of three available layouts. Choose from our vast selection of experimentally validated LNA miRNA PCR primer sets and reference gene primer sets for human, mouse and rat miRNA quantification.
  • 384-well, ready-to-use: design your panels from one of four available layouts. Choose from our vast selection of experimentally validated LNA miRNA PCR primer sets and reference gene primer sets for human, mouse and rat miRNA quantification.

Custom miRNA PCR Panels come with a pre-positioned interplate calibrator (IPC) in replicates that allow optimal calibration and control assessment in multi-plate studies. The IPC is mandatory but can be re-positioned to suit your preferred plate layout. In additional, the panel is displayed with one or more pre-positioned UniSp6 RNA Spike-in controls (CP). RNA spike-in controls may be used for monitoring the cDNA synthesis or RNA purification. The UniSp6 CP assay can be removed and substituted by miRNA primer set(s) or other RNA spike-in control assays. Alternative RNA spike-in controls include UniSp2, UniSp4, UniSp5 and cel-miR-39-3p.

Add custom assays
It is now possible to add custom assays to your custom plate. Custom assays are designed through one of our custom assay design tools. Design LNA-enhanced PCR primers for sequences not available among our predesigned assays, e.g., for novel miRNAs identified using next-generation sequencing.
Procedure
miRNA expression profiling with miRCURY LNA miRNA PCR Panels is straightforward and robust. First, prepare cDNA using the miRCURY LNA RT Kit. Second, add a premix of cDNA, miRCURY LNA SYBR Green PCR Master Mix and RNase-free water to a miRCURY miRNA PCR Panel. Third, run the reaction in a real-time PCR cycler.
Applications
miRCURY LNA Custom PCR Panels are used as part of the miRCURY LNA miRNA PCR System for:
  • Pathway- and disease-focused miRNA profiling
  • Biomarker discovery
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