With the increase in probe length, the quenching efficiency of single-quenched probes diminishes, while the emergence of double-quenched probes effectively addresses this issue. Conventional probes carry a fluorescent moiety at the 5' end and a quencher at the 3' end. SBS Genetech's introduction of double-quenched probes involves the addition of an extra quencher moiety within the conventional probe, reducing fluorescence "leakage". This enhancement enables the attainment of lower fluorescent background signals in experiments, thereby enhancing detection sensitivity.

Advantages

The utilization of high-quality fluorescent probes can significantly enhance assay efficiency and detection accuracy. Evaluation criteria for these probes primarily encompass selectivity, sensitivity, response speed, fluorescence enhancement factor, stability, and biocompatibility. Notably, conventional nucleic acid probes exhibit only one type of fluorescence quenching effect! The effectiveness of the quencher moiety in extinguishing fluorescence relies on distance; proximity results in efficient quenching, whereas greater separation leads to diminished quenching and heightened background fluorescence signals. Double-quenched probes offer a compelling solution to this "distance" challenge, thereby markedly improving qPCR assays!

• The dual-quenching mechanism effectively reduces background signals, enhancing sensitivity.
• It reduces fluorescence signal crosstalk in multi-channel detection, ensuring compatibility.
• Probe lengths can be designed up to 40 nucleotides, offering improved selectivity.

Comparison

Note:

• "√" denotes the presence of the feature.
• "√√" indicates a higher level or enhanced feature compared to the standard probe.