TOCE™ Technology is very elegant oligonucleotide chemistry solution for the homogenous high multiplex real-time PCR. TOCE™ technology overcomes the current technical limitations of target-based probe real-time PCR technology and fully exploits the potential of real-time PCR in high multiplex analysis, through unique signal generation & melting temperature analysis by novel components. TOCE™ technology enables to detect and differentiate multiple targets in a single tube and also can provide quantitative result by cyclic-CMTA analysis.

The new paradigm for High Multiplex real-time PCR

  • High multiplicity in a single channel using Catcher-Tm analysis
  • Controllable Catcher-Tm profile
  • Consistent Catcher-Tm values regardless of target sequence variations
  • As sensitive as singleplex real-time PCR
  • Multiple quantitative analysis using cyclic-CMTA in a single channel
  • High multiple point mutation detections in a single tube

Principles of TOCE™ technology

The key components for TOCE™ technology are DPO™ primer pairs, Pitchers and Catchers. The DPO™ provides highly specific amplification of the target region. The Pitcher is a tagging oligonucleotide which hybridizes specifically to the target region. The Catcher is a dual-labeled artificial template.

The 5’ nuclease activity specifically cleaves a target-bound Pitcher and then designed tagging portion is released. The released tagging portion hybridizes to the capturing portion of the Catcher. The duplex catcher formation induces extension on the Catcher, thus results in the generation of fluorescence signal. The signal can be analyzed in manners of real-time or melting curve analysis.

Quantitative analysis by cyclic-CMTA

In the left panel, CMTA points, preselected cycles during the amplification process where melting temperature analysis is performed, are indicated (①,②,③). The right panel shows the appearance of the melting peak for three different titers of the target. The melting peak appears at the first CMTA point for the high titer, at the second CMTA point for the intermediate titer, and at the third CMTA point for the low titer.
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