What is MGB quencher?

What is MGB quencher?

Nonfluorescent quencher (NFQ) maximizes sensitivity TaqMan MGB probes incorporate an NFQ to absorb (quench) signal from the fluorescent dye label at the other end of the probe. The properties of the NFQ combined with the short length of the MGB probe result in lower background signal than with non-MGB NFQ probes.

What is a minor groove binder?

Minor Groove Binders or MGBs. Minor Groove Binders or MGBs are crescent-shaped molecules that selectively bind non-covalently to the minor groove of DNA, a shallow furrow in the DNA helix. Binding to DNA with specific sequences usually takes place by a combination of directed hydrogen bonding to base pair edges.

How do MGB probes work?

MGB Probes for gene expression analysis & SNP genotyping The double-stranded DNA is denatured by raising the temperature. At this point the fluorescent dye of the probe is quenched by the MGB-Eclipse quencher. By decreasing the temperature the PCR primers and MGB probe anneal to their specific target sequence.

How much does MGB increase TM?

The hydrophobic binding of the MGB to the DNA helix increases the Tm by 15°C–30°C, which al- lows the use of shorter probes with high specificity (10).

What is a non fluorescent quencher?

Q20132 (SE) Essentially nonfluorescent quencher. Long-wavelength absorption. Efficient energy transfer acceptor from red- and near-infrared–fluorescent dyes in FRET applications.

What is the major groove and minor groove of DNA?

The major groove occurs where the backbones are far apart, the minor groove occurs where they are close together. The grooves twist around the molecule on opposite sides. Certain proteins bind to DNA to alter its structure or to regulate transcription (copying DNA to RNA) or replication (copying DNA to DNA).

What is the minor groove of DNA?

The minor groove contains the pyrimidine O-2 and the purine N-3 of the base pair, and the major groove is on the opposite side of the pair. The methyl group of thymine also lies in the major groove. In B-DNA, the major groove is wider (12 versus 6 Å) and deeper (8.5 versus 7.5 Å) than the minor groove (Figure 27.8).

What is the difference between TaqMan and SYBR Green?

In quantitative gene expression analysis, two methods have more popularity, SYBR Green and TaqMan, SYBR Green is relatively cost benefit and easy to use and technically based on binding the fluorescent dye to double-stranded deoxyribonucleic acid (dsDNA) where TaqMan method has more expensive and based on dual labeled …

How long are TaqMan probes good for?

12 months
How long are my custom TaqMan primers or probes stable for? Custom TaqMan primers and probes are good for at least 12 months from the date of receipt if stored properly.

What is TaqMan qPCR?

TaqMan probes are hydrolysis probes that are designed to increase the specificity of quantitative PCR. The TaqMan probe principle relies on the 5´–3´ exonuclease activity of Taq polymerase to cleave a dual-labeled probe during hybridization to the complementary target sequence and fluorophore-based detection.

What is TaqMan QPCR?

Why are minor groove binders used in quenchers?

Because the NFQ does not fluoresce, it produces lower background signals, resulting in improved precision in quantification. The minor groove binder moiety (MGB) increases the melting temperature (T m) without increasing probe length. It also allows the design of shorter probes.

How does a minor groove binder bind DNA?

… Minor Groove Binders or MGBs. Minor Groove Binders or MGBs are crescent-shaped molecules that selectively bind non-covalently to the minor groove of DNA, a shallow furrow in the DNA helix. Binding to DNA with specific sequences usually takes place by a combination of directed hydrogen bonding to base pair edges.

Which is a conjugated minor groove binder ( MGB )?

These fluorogenic probes, known commercially as MGB Eclipseprobes, are characterized by having a conjugated minor groove binder (MGB) ligand at the 5′-end and a fluorophore at the 3′-end. Additionally, they have an efficient quencher moiety at the 5′- end that is useful with a wide variety of fluorescent dyes.

How are nonfluorescent quenchers used in TaqMan MGB probes?

… Molecules that are attached to the 3′ end of TaqMan® MGB probes. When the probe is intact, the nonfluorescent quencher (NFQ) prevents the reporter dye from emitting fluorescence signal. Because the NFQ does not fluoresce, it produces lower background signals, resulting in improved precision in quantification.