metabion´s successful efforts to improve and enhance probe-based real-time PCR assay performance allow us to now offer our HP DQPs!

High Performance Double Quenched Probes have been developed to advance assay accuracy by

• increased probe Tm
• increased thermostability and annealing efficiency
• increased specificity and sensitivity

not only but also for longer probes (>25 nts).

HP DQPs provide consistently reduced Cq values and improved sensitivity, and thus offer solutions specifically for challenging real-time quantitative PCR applications including detection of low-abundance targets or setting up complex multiplex assays.

Our HP DQPs utilize a high performance internal abMFQ quencher strategically positioned between the fluorophore and the 3' MFQ quencher.

In 5' nuclease assays, the probe length is essential to balance quenching efficiency and binding stability. While shorter probes are ideal, certain assays, such as AT-rich targets, may require a longer sequence for optimal Tm tuning. In this case, an internal quencher helps to extending the probe usability beyond the 30-nt window.

For probes longer than 25nts, the internal quencher of a DQP is typically placed between the 9th and 10th nucleotide downstream the 5´-fluorescent reporter. 
 

Thereby the distance between reporter and quencher is shortened, and background/baseline fluorescence is substantially reduced — up to four times as compared to traditional single-quenched probes.

This improvement leads to clearer signals (increased Δ) and thus improved accuracy in your experimental results.

Figure 1. (A) Signal increase of a 28mer HP DQP (FAM-abMFQ-MFQ, green) compared to the single quenched FAM-BHQ®-1 equivalent (pink). (B) Starting fluorescence levels of single-quenched FAM-BHQ®-1 probe (pink) and HP DQPs (FAM-abMFQ-MFQ, green).

However, our HP DQPs not only increase thermostability but also enhance annealing efficiency between probe and target sequence. Advanced kinetics with regard to probe-target hybridization combined with increased thermostability of the probe-target hybrid are leading to a significant increase in signal intensity of up to 80% compared to single-quenched probes.

This effect leading to a substantially improved diagnostic detection sensitivity is also seen for “regular length” probes (18-25nts). Replacing single-quenched dlps by HP dqps may therefore help to improve assay performance for challenging single- and multiplex applications.

metabion's HP DQPs standard portfolio offers an exhaustive range of reporter-quencher combinations. We welcome customization beyond the standard offer and invite you to discuss your special requirements.

Our HP DQPs are available in standard yield ranges, from a minimum of 5 nmol up to 70nmol. Should you require larger amounts of dqp material, we are again happy to discuss your requirements.

Explore our HP (High Performance) Double Quenched Probes and let us support you in reaching next-level diagnostic assay accuracy and sensitivity.

For more information on customizing your probe design contact us at info@metabion.com!

Our standard High Performance Double Quenched Probes - Portfolio

5 yield ranges based on independent lengths of delivered oligonucleotide quantities from 18 to 40 base lengths:

• ≥   5 < 10 nmol
• ≥ 10 < 20 nmol
• ≥ 20 < 30 nmol
• ≥ 30 < 50 nmol
• ≥ 50 < 70 nmol

For ranges ≥ 70 nmol, please inquire.

All probes are HPLC purified and Mass-checked by default!

Explore our offerings today to experience the difference in assay performance! Order here.
 

For your convenience, the High-Performance Double-Quenched Probes (HP DQPs) Flyer is available for download in our Downloads section.