BHQ™ Probes are traditional, linear, dual-labelled FRET probes, typically 20 to 30 bases in length, with a fluorophore and quencher covalently attached to the 5' and 3' ends, respectively. Fluorescence is released through the 5' exonuclease activity of Taq polymerase, which cleaves off the fluorescent dye upon the probe's hybridisation to its complementary sequence. BHQ Probes are ideal for detecting the presence and quantifying the amount of specific target sequences.
Key benefits:
- Unmatched quality and technical support from the original makers of the BHQ quenchers
- Complete suite of fluorescent dyes spanning the entire visible spectrum
- Simple probe design and implementation with consistent and reliable performance
Design versatility without performance variability
When it comes to assay design, we don’t believe in a one-dye-fits-all strategy. Our wide selection of dyes empowers you to design the best PCR or qPCR probe for your application without sacrificing performance. We have paired each dye with the BHQ quencher that offers optimal quenching efficiency. With our versatile BHQ Probes, you can design with confidence.
|
5' fluorescent dye |
Abs (nm) |
Em (nm) |
3' quencher |
|
|---|---|---|---|---|
| FAM | 495 | 520 | BHQ-1 | |
| TET | 521 | 536 | BHQ-1 | |
| CAL Fluor Gold 540 | 522 | 544 | BHQ-1 | |
| CIV-550 | 530 | 550 | BHQ-1*, BHQ-2 | |
| JOE | 529 | 555 | BHQ-1 | |
| HEX | 535 | 556 | BHQ-1 | |
| CAL Fluor Orange 560 | 538 | 559 | BHQ-1 | |
| Quasar 570 | 548 | 566 | BHQ-2 | |
| Cy3 | 549 | 566 | BHQ-2 | |
| TAMRA | 557 | 583 | BHQ-2 | |
| CAL Fluor Red 590 | 569 | 591 | BHQ-2 | |
| ROX | 586 | 610 | BHQ-2 | |
| CAL Fluor Red 610 | 590 | 610 | BHQ-2 | |
| CAL Fluor Red 635 | 618 | 637 | BHQ-2 | |
| Cy5 | 646 | 669 | BHQ-2 | |
| Quasar 670 | 647 | 670 | BHQ-2*, BHQ-3 | |
| Quasar 705 | 690 | 705 | BHQ-2*, BHQ-3 | |
* Recommended quencher
Use the spectral overlay tool to view compatible dyes for your instrument. Learn more
High quality FAM/BHQ-1 ValuProbes at a low price
Our ValuProbe is comprised of 10 nmols probe with a 5' FAM and a 3' BHQ-1 quencher, delivered dried down, purified via Reverse Phase HPLC, and verified via mass spectrometry and UHPLC.
How to order a ValuProbe
- Navigate to the Order now tab at the top of this page.
- Select valuprobe to populate ValuProbe parameters.
- Enter your probe name and sequence.
- Add primers (optional).
- Add to basket.
A ValuMix assay built exactly to your specifications
BHQ Probes in the ValuMix assay format for qPCR and gene expression include a BHQ Probe and two primers mixed at your designated primer to probe ratio between 1 and 4.5. Simplify and streamline your experimental setup while reducing any chance of pipetting error.
Key features of ValuMix assays
- Probes purified via RP HPLC
- Salt-free primers
- Wide selection of dyes: FAM, TET, CAL Fluor Gold 540, CIV-550, HEX, CAL Fluor Orange 560, CAL Fluor Red 610 and Quasar 670
- 0.5 nmol (FAM only), 5 nmol, or 20 nmol delivered
Design considerations for BHQ Probe
- Target a probe length typically between 20 and 30 bases.
- If your sequence is shorter than 17 bases, consider ordering this sequence as a BHQplus Probe.
- BHQ Probe sequences longer than 30 bases are unlikely to have efficient quenching. Consider ordering a BHQnova for sequences 30 bases or longer. For assistance, please contact us.
- Target a %GC content between 30% and 80%.
- Avoid runs of identical nucleotides.
- Do not place a guanosine on the 5′ end. A guanosine next to the reporter dye will alter the fluorescence.
- Avoid guanosine on the 3′ end (ex. 5′-...GGG-3′ or 5′-...GGAG-3′).
- Avoid 4 consecutive guanosines as these can form a stable secondary structure.
- For FAM-labelled probes, avoid a guanosine in the second position on the 5′ end.
- Building probes with an internal quencher (currently available with FAM only) introduces an additional T base into the sequence.
- Use IUPAC code when entering wobble/degenerate bases in the Sequence Entry field instead of using parentheses, e.g. (A/G), (A/G/C/T), etc.
