Integrated DNA Technologies this week released a double-quenched probe that the company said increases the accuracy and reliability of 5’ nuclease qPCR experiments.
As opposed to traditional probes, which have 20 to 30 bases between the dye and quencher, IDT's probe design positions an internal ZEN quencher nine bases from the 5’ fluorophore. "This shortened distance, particularly when combined with the standard 3’ quencher, significantly decreases background fluorescence and increases sensitivity," the company said.
IDT added that the chemical structure of the ZEN quencher stabilizes duplex formation, which allows it to be used in previously validated sequences.
The company said that the double-quenched probes maintain a consistently low background at 40 base pairs or longer, which results in lower Cq values when compared to traditional probes and leads to an increase in specificity without any loss in sensitivity or quenching efficiency.