简介：

BHQ-2 amine is a dark quencher that c beconjugated to target substrates or biomolecules containing a carboxylatemoiety. The BHQ-2 amine is for solution phase conjugations only. We do notrecommend using this product for labeling of peptides d proteins

外观：黑紫色粉末

Absorption Maximum (Lambda Max): 579nm

Storage Conditions: +2 to +8 °C

核磁图谱：

TCL点板纯度测试：

其他介绍：

Two commonly used quenchers, TAMRA™ d DABYCL, limit the ultimatesensitivity d flexibility of qPCR. TAMRA is not a dark quencher d thereforecontributes to overall increase in background because of its own nativefluorescence. DABCYL, though a dark quencher, has inadequate absorptionfootprint that overlaps very poorly with reporter dyes emitting above 480 nm (Figure1).

Figure 1. Absorption spectra of DABCYL-T9with the emission spectra of the commonly-used reporters 6-FAM™, TET™, dJOE™. DABCYL was normalized at the poly-T absorbce (260 nm). RFU = relativefluorescence unit.

Though DABYCL works well in Molecular Beacons (structured probes withpaired reporter d quencher that utilize static quenching), its absorptionmaximum of 474 nm places it below the maxima of the reporters shown in Figure1, therefore it is a poor choice forDual-Labeled Probes (linear probes with separated reporter d quencher thatutilize FRET quenching).

Black Hole Quencher 1 (BHQ^®-1) (with absorption maximum of534 nm) is absorbed at higher wavelengths th DABCYL d is directlysuperimposable with the emission maxima of FAM, TET d JOE. This provides asignifict increase in both static d FRET quenching efficiencies, whichmakes it excellent quencher for Molecular Beacons d Dual-Labeled Probes.As shown in Figure 2, BHQ-1, BHQ-2, d BHQ-3 cover thespectrum from 480 nm into the near IR making it possible to utilize reportersthat emit ywhere in this rge.

Figure 2. Absorption spectra of the threeBHQ varits (conjugated to T-9 d normalized to the poly-T absorbce of 260nm) with the emission maxima of the commonly-used reporters 6-FAM, TET, HEX™,Cyine 3, TAMRA, ROX™, LC Red 640, d Cyine 5.

In addition to better spectral overlap, BHQ probes have much highersignal-to-noise ratios when compared to the corresponding DABCYL or TAMRAprobes (Figure 3).

Figure 3. Signal-to-noise (S:N) ratios werecalculated by dividing the fluorescence signal of a 25mer in the presence of afive-fold excess of exact complementary target sequence by the fluorescenceintensity of the probe alone.

Finally, as shown in Figure 4,the BHQ varits readily permit single-tube multiplexing due to the increasedvariety of reporters that c be effectively quenched with little or nocross-talk. This simplifies the design, implementation, d interpretation ofmultiplexed assays.

Figure 4. The unique characteristics of theBHQ varits permit flexibility in the choice of spectrally well-resolvedreporters, which enable single-tube multiplexing with little or no cross-talk.

In summary, Black Hole Quencher:

1.      has no native fluorescence(emits heat instead of light), resulting in lower background fluorescence

2.      has increasedsignal-to-noise ratio, providing higher sensitivity

3.      enables a wider choice ofreporters for multiplexing