MGB-Eclipse Probes for use in 5′ nuclease real-time PCR assays 

MGB Probes provided by Eurofins Genomics are licensed under MGB Eclipse patents for use in human IVD applications. These MGB Probes contain a 5’ fluorescent dye and the 3’ Eclipse quencher (EQ) along with the minor groove binder (MGB) molecule which is also linked to the 3' terminus.

Ordering

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Dye-quencher combinations

Eurofins Genomics' MGB Probes are licensed for human in vitro diagnostics

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Pricing

Pricing is located on the price list page, under modifications.

Specifications & Details

Product specifications*:

• Quantity delivered: 5, 20 and 40 nmole
• Length of the custom DNA sequence: 5–40 bases
• Supplied after QC by MS and purification by HPLC
• Delivery Time: 6–8 working days
• Delivery format: lyophilized in tubes

A ready-to-use qPCR probe dilution buffer (10 mM Tris-HCl; 1 mM EDTA; pH 8) is provided along with all MGB Probes (1 mL for up to 3 probes).

Eurofins Genomics' MGB Probes are licensed for human in vitro diagnostics applications to provide information for individual patient management solely using 5' nuclease (Taqman-MGB) technology for research grade licensed products and IVD grade licensed products. MGB Eclipse is a registered trademark of Elitech Group.

MGB molecule in real-time PCR
A probe containing a MGB molecule forms an extremely stable duplex, resulting in a significant increase in its melting temperature (T_m). Therefore, short probes (down to 13 bases) can be used in real-time PCR. Two prominent use of MGB probes are for gene expression & SNP genotyping

Gene Expression analysis:
The annealing of the double-stranded DNA with the MGB probe and other primers leads to the formation of duplexes. At this point the fluorescence of the dye at the 5'-terminus of the MGB probe is quenched by the MGB-Eclipse quencher.

Figure 1: Gene Expression analysis

Primer extensions of the forward and reverse primers are carried out by the taq polymerase. When the taq polymerase encounters the MGB probe, its 5' nuclease activity leads to the cleavage of the probe and, consequently, the separation of the fluorescent dye into the solution. Since the dye in solution is not in the immediate proximity of the quencher, significant enhancements in the unquenched emission signal are observed. The levels of the target DNA can be measured by monitoring the enhancements in the flourescent signal with each new PCR cycle.

Allelic discrimination by selective annealing

Single nucleotide polymorphisms, also called SNPs (or “snips”), are a common type of genetic variation. Each SNP represents a difference in a single DNA base within a sequence—e.g., replacement of a 'G' witgh a 'T'. MGB probes can also be used to identify such SNPs or to identify the levels of different alleles within the strech of DNA. Allelic discrimination is achieved by the selective annealing of the custom sequences of the MGB probes.

Figure 2: SNP genotyping

Additional Probe Options