QuantiNova LNA PCR Custom Assay (200)

Custom LNA-enhanced assays from QIAGEN provide accurate and sensitive gene expression analysis using EvaGreen-based digital PCR. The QuantiNova LNA PCR Custom Assays support any RNA target and offer full Ensembl database support for human, mouse and rat targets, streamlining assay creation. They are well-suited for targets lacking predesigned assays, for discriminating splice variants, and for verifying novel mRNAs or lncRNAs. For digital PCR, the assays have been optimized with the QIAcuity EG PCR Kits and the QIAcuity instrument, creating a simple and fast absolute quantification workflow that takes only 2 hours. After cDNA synthesis using the QuantiTect Reverse Transcription Kit, digital PCR provides highly precise target quantification, detecting even the smallest expression changes at the lowest concentrations. A subset of key assays have been experimentally validated on the QIAcuity instrument. The products are intended for molecular biology applications and are not intended for the diagnosis, prevention, or treatment of a disease.

Stringent design criteria and lab-validated algorithms deliver high specificity and efficiency. The high binding affinity of LNA bases increases primer placement flexibility, enabling better target discrimination and robust quantification for AU-rich targets, low-abundance transcripts, targets with high secondary structure, and highly complex samples. Sensitivity ensures excellent amplification efficiencies down to 1 RNA molecule, and clever placement of LNA provides a high signal-to-noise ratio, allowing discrimination of sequences that differ by only a single nucleotide and eliminating non-specific amplification and primer-dimer formation. The sophisticated LNA design algorithm incorporates over 50 different parameters, all thoroughly lab-validated, and has been used to design over 1.3 million assays. The EvaGreen-based QIAcuity EG PCR Kit uses an intercalating dye that binds double-stranded DNA, and the dPCR nanoplate is divided into thousands of partitions, so the primer concentration needed for the dPCR reaction is only half that required for qPCR.

The custom assay design tool supports human, mouse and rat mRNA and lncRNA targets of at least 55 nucleotides, blasting against species-relevant databases including Ensembl and RefSeq. Known gene targets are designed as intron-spanning by default to prevent gDNA amplification. Options exist for transcript-specific designs to discriminate splice variants, SNPs or isoforms. Users can submit up to 10 different target sequences at once. Functionally validated human, mouse and rat Reference Gene Assays are available for normalization. The QIAcuity Software Suite is used for analysing dPCR data, providing results as fold change and fold regulation. Assays are shipped at ambient temperatures, and in-stock assays are delivered within 1–5 days.