The advanced algorithm evaluates approximately 3,000 primer pair designs based on more than 60 different criteria to find LNA optimal primer sets for your miRNA within a few minutes. The miRCURY LNA miRNA Custom PCR Assay design tool lets you easily design highly sensitive and specific LNA-enhanced PCR primer sets for any miRNA not available as a predesigned product. T m-normalization and LNA enhancement give the primers a higher binding affinity than standard DNA primers, dramatically increasing assay sensitivity and specificity.Ĭompared with other miRNA PCR systems that use either stem-loop or standard DNA primers, the LNA-enhanced primers offer significantly increased sensitivity, especially for AT-rich miRNAs Principle The rigorous assay selection process ensures that each primer set delivers the highest specificity and efficiency for the most reliable and accurate results. MiRCURY LNA miRNA PCR Assays have been developed using stringent design criteria and lab-validated algorithms. A subset of key miRNAs have been experimentally validated on the QIAcuity instrument. After cDNA synthesis using the miRCURY LNA RT Kit, digital PCR provides highly precise target quantification, detecting even the smallest expression changes at the lowest concentrations. This can be quantified by two PCR methods: qPCR with standard real-time thermocyclers and digital PCR using the QIAcuity instrument and the QIAcuity EG PCR Kit. MiRCURY LNA miRNA PCR Assays were developed to provide high-specificity target amplification. TAMRA is also a quencher option for a FAM reporter dye.Optimized digital PCR analysis with the QIAcuity EG PCR Kits and the QIAcuity instrument We supply commonly used dark quenchers as well as the proprietary dark quenchers, Iowa Black FQ, Iowa Black RQ, and the internal ZEN Quencher. Double-quenched probes are available with a variety of fluorophores, including SUN, FAM, HEX, TET, Yakima Yellow, and Cy 5. These double-quenched probes are an improvement over traditional dual-labeled probes and have consistently low background, reduced C q values, improved precision, and enable the use of longer probes for design in AT-rich regions. Dark quenchersĪre compatible with a broad range of image analysis instruments. This characteristic allows for expanded options for multiplex assays. Quenchers: Traditional dark quenchers absorb broadly and do not emit light, which allows for the use of multiple reporter dyes with a given quencher.As an additional reference, for a list of selected IDT dyes and quenchers, please see the dye and quencher wavelength figure and the instrument compatibility table. Which helps you select dyes based on what instrument you have. For assistance in selecting multiplex dye combinations, use our Multiplexing Dye Selection Tool, For multiplexing applications, we recommend using reporter dyes with the least amount of spectral overlap. Please see the instrument compatibility table for a list of reporter dyes compatible Reporter dyes: The correct reporter dye will depend on the type of instrument you are using and the compatibility of the dye with the instrument.Selecting the correct reporter dye and quencher
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