PlexPCR™ utilises the PlexZyme™ technology to offer high performance and reliable qPCR detection. During PlexPCR™ reaction, primers amplify target nucleic acid sequences and produce amplicons which serve as a template for PlexZyme™ formation. Once the partzymes have assembled into PlexZymes™, universal probes bind and enzymatic cleavage of the probes between fluorophore and quencher dye pairs leads to an increase in fluorescence. Changes in fluorescence allow detection and/or quantification of the target nucleic acid in real time.
- Can detect < 10 copies of target
- Good dynamic range from 106 to 10 copies
- Specificity is determined by 4 binding events of the 2 primers and 2 partzymes to the amplicons
- New targets can be linked to existing probes simply by changing the sequences of the sensor arms of the partzymes.
PlexPCR™ 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 flexible 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 probes.
Principle Of Multiplexing With PlexPCR™
In a multiplex reaction, the universal probes are labelled with different fluorophores so that fluorescence signal corresponding to detection of each target sequence can be monitored simultaneously in real time. The highly multiplex nature of PlexPCR™ can maximise the outputs of qPCR instruments.
New Benchmark In Multiplexing – 20 Target Multiplexing With PlexPCR™
Simultaneous amplification of 20 genes showing robust, efficient and sensitive multiplexing down to 30 copies. The reactions contained 20 primer sets, 20 Plex
Zymes™, buffer, dNTPs and Taq in one mix read by 20 universal probes in 4 wells. Human genomic target concentration used in the reaction: 100 ng, 25 ng, 6.3 ng, 1.6 ng, 391 pg, 98 pg and no DNA.
Mokany E., Bone S.M., Young P.E., Doan T.M., Todd A.V., 2010. MNAzymes are a versatile new class of nucleic acid enzymes which can function as biosensors and molecular switches. J. Am. Chem. Soc. 132(3); 1051-1059
Link | Article | Supplementary Information