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Technologien
TripleHyb Technology - a new real-time detection method
The patent pending TRIPLEHYB technology is a novel detection format for real-time PCR. This format besides a conventional set of primers comprises a pair of labeled oligonucleotide probes. Both probes consist of a target-complementary subsequence and a target-unrelated subsequence which has the capability to form a stem structure between the probes. In the presence of target the upstream probe hybridizes by its target-complementary 5` end and the downstream probe adjacently with its 3` target-complementary end to the desired sequence. Simultaneously, an intermolecular stem structure is formed between the 3` end of the upstream and the 5` end of the downstream probe thus stabilizing the structure of the detection complex. A Taq polymerase which possesses 5` 3` exonuclease activity cleaves the upstream probe labeled with both a reporter dye and quencher dye, respectively. The dyes are separated upon cleavage, resulting in an increase in fluorescence for the reporter. Thus, the increase in fluorescence is directly proportional to the target amplification during PCR. TRIPLEHYB is a powerful alternative to all known real-time PCR detection platforms regarding target quantification and particularly genetic polymorphism analysis.
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