Cycleave PCR™ technology

Cycleave PCR combines the use of Ex-Taq R-PCR with Cycling Probe Technology*. The later permits particularly sensitive and specific detection of amplificon.
It is described in the figure below.

Principle of cycling probe technology (CPT)

Using this technology Takara developped several kits:
Cycleave PCR kit series:
o CycleavePCR Bacillus anthracis Detection Kit Ver.1.1
o Bovine Sexing Kit
o Mycoplasma pneumoniae Detection Kit
o E.coli O157 (VT1/VT2) Detection Kit
o Salmonella Detection Kit
o Meat Species Identification Kit
These kits use multiplex PCR to compare amplification of the target with an internal control or to detect different targets in the same tube.

Cycleave kits on demand
Equivalent kits can be developed by TaKaRa under specific customer requirements. Please inquire by e-mail!

CPT*: technology description

Cycling probe is a chimeric RNA-DNA probe that hybridize to a target sequence of amplified gene. Once hybriydized to amplification product, RNA part of the probe is cleaved by RNase H. Actually this enzyme recognize specifically RNA-DNA duplex. Thus Fluorescer and Quencher on each side of the probe are separated and fluorescence is emitted. This fluorescence increases whith the number of amplified DNA molecules allowing to detect specific sequences. Measurement of the fluorescent intensity, which increases proportionally to the rate of probe cleavage, allows monitoring of the amount of amplification products. This provides rapid results without the need for electrophoresis. Use of multiple probes with different dyes together with an apparatus able to detect diferent fluorescer, allows to have an internal control amplified in the same time as the target. With probes differeing by a single nucleotide in its RNA part it is possible to detect SNP (Single Nucleotide Polymorphism). Also multiplex PCR with more than one primer set and probe can detect different genes in the same tube.

* Sold under license from ID Biomedical Corp.