Transcript PPT

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
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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.
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Real Time/Quantitative PCR
Black Hole Quencher Dyes
The Inescapable SolutionSM for DNA Hybridization Probes
BHQ Dye Absorption Maxima and Quenching Range
Quencher
BHQ-1
BHQ-2
BHQ-3
Abs (nm)
Quenching Range (nm)
534
579
672
480-580
550-650
620-730
BHQ Dye / Reporter Combinations
The unique characteristics of the BHQ
classof quenchers permits flexibility in the
c h o i c e o f s p e c t r a l l y w e l l - r e s o l ve d
fluorophores enabling single-tube
multiplexing with little or no cross-talk
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Quencher
Suggested Fluorophores
BHQ-1
BHQ-2
BHQ-3
FAM, TET, JOE, HEX, …
TAMRA, ROX, CY3, LCRed 640, …
CY5.5, IRDye700, LCRed 705, …
Real Time/Quantitative PCR
Black Hole Quencher Dyes
The Inescapable SolutionSM for DNA Hybridization Probes
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.
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