MNAzyme qPCR

During MNAzyme quantitative PCR (qPCR), primers amplify target nucleic acid sequences and produce amplicons which serve as templates for MNAzyme formation. Once the partzymes have assembled into MNAzymes, universal reporter probes bind and enzymatic cleavage of the probes between fluorophore and quencher dye pairs leads to increases in fluorescence. Changes in fluorescence allow detection and/or quantification of the target nucleic acid in real-time.

Multiplex MNAzyme qPCR

MNAzyme qPCR has many attributes which make it superior for multiplexing. A series of well characterized universal reporter probes can be incorporated into multiplex assays allowing analysis of several targets simultaneously. The modular nature of the partzymes means that new targets can be linked to existing probes simply by synthesising new partzymes with sensor arms tailored to the new target, and substrate arms to one of the series of unique universal reporter probes.

Advantages of MNAzyme qPCR

Universal reporter probe

Use any universal reporter probe to detect any specific target
Use any universal reporter probe for any application (quantification, methylation analysis etc)
Obtain reliable, consistent performance using well characterised probes which behave in a predictable manner
Reduce synthesis & QC costs by manufacturing dual-labelled probes in bulk
Use the same universal reporter probe for multiple targets
Eliminates probe waste by using residual probe from one assay for your next assay

Multiple unique universal reporter probes

Allow rapid multiplex assay development
Require fewer reactions & less reagents (eg polymerase) & consumables
Obtain more information per specimen, less precious sample used, more replicates can be tested
Allow greater confidence in data as internal controls can be included

Target-specific interrogation by partzymes

Results in assays with greater specificity as a positive signal requires binding of 2 partzymes & 2 primers which results in 4 determinants of specificity
Target-specific MNAzyme formation requires 2 partzymes to recognize 2 complementary sequences that are immediately adjacent on the amplicon
Partzymes are target-specific, but since they are usually comprised of DNA only and are not labeled, they are significantly cheaper to synthesise than target-specific dual labeled probes.

MNAzyme qPCR is the only method which combines
target specific interrogation and signal generation using universal reporter probes