1. Real Time/Quantitative PCR
Competence and Service in Molecular Biology

2. Real Time/Quantitative PCR
metabion offers Black Hole series of dark quenchers (BHQTM) which were designed for use in real-time PCR assays. These molecules function as efficient quenchers of fluorescence over the entire visible spectrum and into the IR, re-emitting their energy as heat rather than light. Probes made with BHQ quenchers exhibit extremely low background noise and allow for lower copy sensitivity. BHQ-1 has its max absorption at 534 nm and quenches most efficently in the 480 to 580 nm range. BHQ-2 has its λmax at 579 nm, which enables it to “absorb” fluorescence in the 550 to 650 range. BHQ-3 has its max absorption at 672 nm with an optimal quenching range of nm.
Competence and Service in Molecular Biology

3. Real Time/Quantitative PCR
BHQ Dye Absorption Maxima and Quenching Range
Quencher Abs (nm) Quenching Range (nm)
BHQ
BHQ
BHQ
BHQ Dye / Reporter Combinations
Quencher Suggested Fluorophores
BHQ FAM, TET, JOE, HEX, …
BHQ TAMRA, ROX, CY3, LCRed 640, …
BHQ-3 CY5.5, IRDye700, LCRed 705, …
The unique characteristics of the BHQ classof quenchers permits flexibility in the choice of spectrally well-resolved fluorophores enabling single-tube multiplexing with little or no cross-talk
Competence and Service in Molecular Biology

4. Real Time/Quantitative PCR
Absorption spectra of DABCYL-T9 and BHQ-1-T9 with the emission spectra of FAM, TET, and JOE. DABCYL and BHQ-1 dye were normalized at the poly-T-absorbance (i.e. 260 nm) to better demonstrate the larger extinction coefficient of the BHQ-1 dye. RFU=relative fluorescence units.
Competence and Service in Molecular Biology