The design process
The miRCURY LNA miRNA Custom Probe PCR Assay design tool lets you easily design highly sensitive and specific LNA-enhanced PCR primer sets and probe for any miRNA not available as a predesigned assay. The advanced algorithm evaluates approximately 3,000 primer pair and probe designs based on more than 60 different criteria to find LNA optimal primers and probe for your miRNA within a few minutes. The tool has been designed for miRNAs but can also be used for other small RNAs 14–27 nucleotides in length.
The design criteria include:
- Optimization of melting temperatures by varying the LNA distribution of the primers and probe to ensure high amplification efficiency and specificity.
- Calculations and adjustments of self-hybridization and cross-hybridization scores for efficient PCR reactions.
- Adjustments to avoid potential primer–dimer formation in the PCR reaction.
- Intelligent positioning of LNA bases in the primers and probe based on our vast knowledge of LNA oligonucleotide design.
- miRBase searches to identify potential miRNAs with high sequence similarity within the same species. This ensures that the primer sets and probe are highly specific for the small RNA for which they were designed.
A unique system for miRNA quantification
The miRCURY LNA miRNA Probe PCR System enables sensitive, specific miRNA quantification and profiling via real-time PCR. Detection is based on a FAM- and dark 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 Custom Probe PCR Assays 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 that uses 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
Optimized and dedicated for the miRCURY LNA miRNA Probe PCR System, this kit contains the high-performance QuantiNova Probe Master Mix and the miRCURY Probe Universal Reverse Primer. Combined with a miRCURY LNA miRNA Probe Assay or a miRCURY LNA miRNA Probe PCR Panel, the chemistry enables miRNA detection via quantitative real-time PCR.
miRCURY LNA miRNA Probe 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 22. The miRCURY Probe Universal Reverse Primer is contained in the miRCURY Probe PCR Kit.
miRCURY LNA miRNA Probe PCR Panels
Ready-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, 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.