Competence and Service in Molecular Biology Real Time/Quantitative PCR.
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Competence and Service in Molecular Biology Real Time/Quantitative PCR Black Hole Q uencher Dyes The Inescapable Solution SM forD N A H ybridization Probes A New C lass ofH igh-Efficiency D ark-Q uencher Real-tim e,quantitative PC R A llelic discrim ination Spectralgenotyping in situ hybridization Single-tube m ultiplexing
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Transcript of Competence and Service in Molecular Biology Real Time/Quantitative PCR.
Competence and Service in Molecular Biology
Real Time/Quantitative PCR
Black Hole Quencher Dyes
The Inescapable SolutionSM for DNA Hybridization Probes
A New Class of High-Efficiency Dark-Quencher Real-time, quantitative PCR Allelic discrimination Spectral genotyping in situ hybridization Single-tube multiplexing
Competence and Service in Molecular Biology
Real Time/Quantitative PCR
Black Hole Quencher Dyes The Inescapable SolutionSM for DNA Hybridization Probes
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 620-730 nm.
Competence and Service in Molecular Biology
Real Time/Quantitative PCR
BHQ Dye Absorption Maxima and Quenching Range
Black Hole Quencher Dyes The Inescapable SolutionSM for DNA Hybridization Probes
Quencher Abs (nm) Quenching Range (nm)
BHQ-1 534 480-580 BHQ-2 579 550-650BHQ-3 672 620-730
BHQ Dye / Reporter Combinations
Quencher Suggested Fluorophores
BHQ-1 FAM, TET, JOE, HEX, …BHQ-2 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
Real Time/Quantitative PCR
Absorption spectra of DABCYL-T9 and BHQ-1-T9with the emission spectra of FAM, TET, and JOE. DABCYL andBHQ-1 dye were normalized at the poly-T-absorbance (i.e. 260nm) to better demonstrate the larger extinction coefficient ofthe BHQ-1 dye. RFU=relative fluorescence units.
Black Hole Quencher Dyes The Inescapable SolutionSM for DNA Hybridization Probes
