miRCURY LNA miRNA Custom Probe Assay

Custom-designed, LNA-optimized miRNA PCR primer sets from QIAGEN enable extremely sensitive and specific probe-based quantification of miRNAs as part of the miRCURY LNA miRNA Probe PCR System. Both the forward PCR amplification primer and the FAM- and dark quencher-labeled probe are miRNA-specific and enhanced with LNA technology, which improves assay sensitivity, specificity and efficiency by allowing shorter primer and probe design while preventing overlap when binding to the target.

Combining universal reverse transcription with LNA-enhanced, Tm-normalized, miRNA-specific primers and probe enables accurate quantification of individual miRNAs from as little as 1 pg total RNA or down to 1 molecule without pre-amplification, with the assays able to detect down to 1 miRNA copy. The chemistry discriminates closely related miRNAs with sequence differences as little as one nucleotide and is well suited for serum or plasma samples and other liquid biopsies such as urine, CSF, exosomes and FFPE samples.

The online design tool evaluates approximately 3,000 primer pair and probe designs based on more than 60 different criteria to identify optimal LNA primers and probe for any miRNA, and can also be used for other small RNAs 14–27 nucleotides in length. The simplified workflow takes 2 hours from RNA to data, with the cDNA generated in the RT step reusable across the miRCURY LNA miRNA Probe PCR System. Robustness allows room temperature setup, and the assays are manufactured under ISO-compliant conditions for high day-to-day and site-to-site reproducibility. The assays are validated with the QuantiNova Probe Master Mix.