Advanced qPCR probes
GeneTAG Technology, Inc.
Conventional dual-labeled qPCR probes lack error-checking mechanisms and cannot provide multi-temperature single base discrimination. GeneTAG Technology has developed 3 error-checking DNA Detection Switch (DDS) probe systems for high-fidelity detection and/or amplification. DDS probes employ the interaction of 2 labeled polynucleotide components: a fluorescent probe and a quenching antiprobe. Probes bind preferentially to their intended targets, turning on signaling, while dissociating from a nearly complementary antiprobe that otherwise turns off signaling. This general signaling mechanism is termed a ′DNA detection switch′.
DDS probes are designed to match the target sequence, and antiprobes are complementary to the probe except for a carefully engineered mismatch. The thermodynamics of this design facilitates exacting target detection because the competitive antiprobes bind and quench free-floating probes, preventing the detection of closely related off-targets. Principles and applications of each error-checking DDS probe system are described below.