Integrated DNA Technologies (IDT) has launched a new double-quenched probe that increases the accuracy and reliability of 5’ nuclease qPCR experiments.
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IDT said that while traditional probes have 20-30 bases between the dye and quencher, the new proprietary probe design positions an internal Zen quencher only 9 bases from the 5’ fluorophore. The shortened distance, particularly when combined with the standard 3’ quencher, decreases background fluorescence and increases sensitivity.
According to IDT the chemical structure of the Zen quencher stabilises duplex formation which allows for its use in previously validated sequences. The improved functionality increases qPCR accuracy and sensitivity when compared to traditional probes.
IDT claimed that due to the incorporation of Zen at a fixed position 9 bases from the 5’ end, the quencher is always within close proximity of the probe. As such, the initial background fluorescence signal is much lower. This makes subsequent changes easily detectable and functionally increases assay sensitivity.
Additionally, while traditional probes do not remain well quenched over 30 base pairs, the double-quenched probes maintain a consistently low background even at 40 base pairs or longer. Thus, double-quenched probes result in lower Cq values when compared to traditional probes. This leads to an increase in specificity without any loss in sensitivity or quenching efficiency. This is particularly useful for targets that are AT-rich and require longer probes.
IDT is a supplier of custom nucleic acids in the US, serving academic, government, and commercial researchers in pharmaceutical, biotechnology, clinical diagnostics.
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