The Fluorescent Effects of BHQ Labeling

The demands for modern genomic and diagnostic applications often focus on the increasing demand for higher detection sensitivity, which has led to the development of a range of new non-fluorescent quenchers. The most commonly used is the Black Hole Quencher reagent (BHQ). Fluorescent quenching generally refers to the process in which the fluorescent molecule reduces the physical or chemical interaction with the solvent or the corresponding solute molecule. The quenching efficiency is improved due to the high extinction coefficient and wide spectral overlap of each BHQ dye. This means that the BHQ dye provides a wider range of wavelength use for different detection purposes, covering the visible light to the near-infrared region (480-730nm). Coupled with the lack of residual background fluorescence in these molecules (a true dark quencher), BHQ dyes are a favorable choice for detection techniques.

Fig.1 The structure of TaqMan probes. (Kostina, et al., 2007)

BHQ Labeling Service at CD BioGlyco

CD BioGlyco provides BHQ dye labeling services, including BHQ-0 (430-520 nm), BHQ-1 (480-580 nm), BHQ-2 (550-650 nm), and BHQ-3 (620-730 nm). Combines the absorption spectra of the four BHQ dyes and covers the range of 430 nm to the near-infrared. The fluorescent wavelengths falling in this range are quenched for all reporter groups, making it suitable for multiplex qPCR.

Among them, BHQ-1 and 2 are our popular labeling products, suitable for 5′ -Phosphoramidites, dT-Phosphoramidites, or 3′ -CPG. If only excitation and emission values are considered, Cy5 requires BHQ-3 or BHQ-2 for Cyanine-5 to quench efficiently.

• BHQ-2 is not easy to degrade and is most effective in quenching fluorescent moieties such as TAMRA and Cy3. BHQ-1 is activated and coupled to substrates with nucleophilic groups, such as amino groups on polypeptides.
• BHQ-1 is located at the ends or in the interior of oligonucleotides and is commonly used in conjunction with fluorescent groups, such as FAM, TET, and HEX.

BHQ quencher is like a black hole and does not produce fluorescence itself. BHQ absorbs all the fluorescence signals emitted by the reporter group within a certain range, solving the problem of light pollution in the qPCR experimental system.

Table 1. Classification of BHQ. (CD BioGlyco)

Applications of BHQ Labeling

• BHQ-labeled oligonucleotide can be used in molecular diagnosis and disease treatment.
• BHQ-labeled oligonucleotide plays a key role in the accumulation of PCR product detection in real-time.
• BHQ-labeled oligonucleotide can be used as part of the double-quenching probes available.

Advantages

• BHQ-labeled oligonucleotide with a broader coverage of the absorption spectrum.
• The BHQ labeling service we provide greatly reduces the fluorescent background signal.
• Our professional team generates Custom Carbohydrate Synthesis services for our clients according to their requirements.

CD BioGlyco provides high-quality, multi-category primer synthesis services promptly to our clients. We are capable of providing efficient and reliable services to accelerate our clients' scientific research by shortening the project time. If you are interested in our project, please feel free to contact us.