PentaBase is offering high quality standard DNA probes and primers but also improved probes and primers. PentaBase offers two different types of dual labeled probes for Real-Time PCR: Molecular Beacons/EasyBeacons™ and TaqMan®/HydrolEasy™ probes. Dual labeled probes normally contain a quencher at the 3'-end and a fluorophore at the 5'-end, but can also contain the fluorophore and/or quencher internally in the sequence.
EasyBeacons™ - PentaBase's alternative to Molecular Beacons
EasyBeacons™ are nuclease resistant probes developed by PentaBase™ to be used in 3-step Real-Time PCR and in situ hybridizations. Like Molecular Beacons EasyBeacons™ are quenched when the probes are unbound, but fluoresce at hybridization to a target sequence. However, unlike Molecular Beacons EasyBeacons™ do not require an internal stem to be quenched efficiently. The pentabases in EasyBeacons™ will, by having hydrofobic interactions with each other in the unbound probe, ensure that the quencher and fluorophore are kept in close proximity and hence resulting in an efficiently quenched probe with low background fluorescence.
Due to EasyBeacons™ nuclease resistance, they can also be used for end-point detection or verification of your real-time signal. This is a valuable feature, especially if you want to discriminate between almost similar genetic sequences like SNPs, alleles or if you want a quality control on your quantification data.
HydrolEasy™ probes - PentaBase's alternative to TaqMan® probes
HydrolEasy™ probes are developed by PentaBase to provide MGB-TaqMan® users and users of all other hydrolysis probes with a probe with significantly better signal-to-noise ratio, a higher specificity and a higher sensitivity than conventional probes. When the HydrolEasy probe is intact the fluorophore and the quencher are, in contrast to "standard" TaqMan® probes, kept in close proximity by hydrophobic interactions between pentabases. During the amplification reaction the probe hybridizes to the target and exonuclease activity of the polymerase cleaves of the fluorophore from the probe whereby a strong fluorescence signal is generated. The figure below shows how a HydrolEasy™ probe degrades in the same way as normal TaqMan® probes do.
