Key featuresShow Hide
- Dark quencher with broad absorbance centred around 650 nm
- A key advantage of the BBQ-650® is its stability to ammonia and oxidation conditions
- Ester for post-synthetic 5' modification of oligonucleotides
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LGC, Biosearch Technologies offers our BlackBerry Quencher 650 (BBQ-650) as a synthesis-stable dark quencher of long wavelength fluorescence. An 8-alkoxyjulolidine moiety was found to be a powerful π-electron donor, affording a surprising bathochromicshift when compared to related compounds. The broad absorbance centred around 650 nm effectively overlaps the emission maxima of popular long-wavelength fluorophores such as Cy™ 3, TAMRA, Texas Red, ROX, Cy™ 5, and Cy5.5, allowing efficient quenching.
Molecular beacon probes bearing various 5'-fluorophores (FAM, Cy™ 3, Texas Red, Cy™ 5, Cy™ 5.5) have been synthesised using 3'-BBQ-650 CPG. Signal-to background ratios upon binding to fully complementary target were noted to be excellent, e.g., >90 with Cy5 and >88 with Cy5.5. The probes are known to be successful in typing C to T transitions at positions 627 and 630 of the human chemokine receptor 5 gene,(1) and produced excellent results in real-time PCR studies.
BlackBerry Quenchers are known to be excellent FRET-mode quenchers. Pairs of complementary strands were designed that would bring a 5'-Cy5.5 fluorophore to within 5 or 10 base pairs (20-40 Å) of a 3'-BBQ-650 upon hybridisation,(2) where quenching efficiencies of ≥98.3 and ≥98.9%, respectively, were observed. Melting temperatures of these hybrids were unchanged from non-labelled hybrids, showing that these quenching efficiencies were due to FRET quenching and not contact quenching.
BlackBerry Quenchers may be installed at the 3’ terminus, internally, or at the 5’ position, using the variety of products available.
- Genotyping SNPs With Molecular Beacons, Marras, S. A. E.; Kramer, F. R.; Tyagi, S. Methods in Molecular Biology 2003, 212, 111-128.
- Efficiencies of fluorescence resonance energy transfer and contact-mediated quenching in oligonucleotide probes. , Marras, S. A. E.; Kramer, F. R.; Tyagi, S. Nucleic Acids Res. 2002, 30, e122.