MGB-Eclipse Probes for 5’ nuclease real-time PCR assays
MGB Probes provided by Eurofins Genomics stabilizes probe-target hybridization and increases melting temperature.
Eurofins Genomics MGB Probes incorporate a 5’ fluorescent dye and the 3’ Eclipse quencher (EQ) attached to the minor groove binder (MGB) molecule.
Product specifications:
- Delivered quantity: 5 nmol, 20 nmol and 40 nmol
- Probe lengths: 5 - 40 bases
- HPLC purified by default
- TAT: 3 – 5 working days
- Delivery format: dried or liquid at selected concentration
- QC report incl. MALDI-TOF MS spectra free of charge
Dye-quencher combinations:
5' Reporter |
Abs [nm] |
Em [nm] |
3' Quencher |
Price-Cat. |
FAM [FAM] |
495 |
520 |
MGB-Eclipse [MGBEQ] |
1 |
Yakima Yellow [YAKYE] |
530 |
549 |
MGB-Eclipse [MGBEQ] |
2 |
HEX [HEX] |
535 |
556 |
MGB-Eclipse [MGBEQ] |
2 |
TexasRed [TxRed] |
583 |
603 |
MGB-Eclipse [MGBEQ] |
1 |
Cyanine5 [CY5] |
647 |
673 |
MGB-Eclipse [MGBEQ] |
2 |
ATTO 647N [ATTO647N] * |
646 |
664 |
MGB-Eclipse [MGBEQ] |
2 |
12MG
Advantage of the MGB molecule in real-time PCR
Probes containing a MGB molecule form an extremely stable duplex which gives the probe a significant increase of the melting temperature (Tm). It is therefore possible to use shorter probes (down to 13 bases) in real-time PCR compared to conventional dual labelled probes.
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. Allelic discrimination is achieved by the selective annealing of the MGB probes (Figure 2).
The taq polymerase synthesises a complementary DNA strand using the PCR primers and template as a guide. When the polymerase reaches the MGB probe its 5´nuclease activity cleaves the probe and separates the fluorescent dye from the non-fluorescent quencher.