Advanced qPCR probes

GeneTAG Technology, Inc.

​Principles of error-checking probes:
​Generic-component DDS probes:
Other DDS probe designs (Universal, Half-Universal, and MacMan probes) use generic components that enable reduced cost and that perform comparably to conventional dual-labeled probes. Follow the links above to learn more about each technology.
G-Force probe principle:
G-Force probes:
Our single-molecule G-Force probes employ guanine-mediated fluorescence quenching by forming a hairpin between adjacent C-rich and G-rich sequences. G-Force probes can also serve as reliable reference probes to provide positive amplicon detection in cases where a second probe is distinguishing a specific mutation, and together, the mutant frequency can be assessed.
Principles of generic-component DDS probes:

GeneTAG offers 7 different DNA Detection Switch (DDS) probe systems that are specialized for high-fidelity detection (error-checking probes) or for low-cost detection (generic-component probes). DDS probes employ 2 interacting components: a fluorescent labeled probe and a nearly complementary quencher-labeled antiprobe. Probes preferentially bind to their intended targets, liberating fluorescence. Specificity is enhanced by providing an excess of antiprobes that intercept any incorrect hybridizations between probes and off-targets, thereby preventing false positives. This general signaling mechanism is termed a ′DNA detection switch′. ​ 
 
Error-checking DDS probes: 
Conventional dual-labeled qPCR probes lack error-checking mechanisms and cannot provide multi-temperature single base discrimination. Our error-checking DDS probes function either as internal probes (iDDS or Flip probes) or primer-probes (ZIPR probes), wherein each target-specific probe is combined with a slightly mismatched, competitive antiprobe. The antiprobe blocks off-target detection over a wide range of annealing temperatures (up to 30°C), and facilitates multiplexing. The internal probes provide reliable single base discrimination and are also useful for validating next generation sequencing results. ZIPR primer-probes are stringent reference probes that enable 1- or 2-color detection of targets of interest, such as gene fusions occurring in cancer. Primer-probes label all amplicons for maximum signaling. Our internal Flip probe system generates linear amplification curves, which are useful for quantitative real-time or endpoint detection. The signaling mechanisms of our error-checking probes are shown below:

Primer-probes

Internal probes

Overview of DNA Detection Switch probe technology