miRCURY LNA miRNA Custom PCR Panels

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).

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.

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.

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