HydrolEasy® Probe

The HydrolEasy® probe is an improved alternative to TaqMan®1 and molecular beacon hydrolysis probes. The INA® technology used in HydrolEasy® probes creates a secondary structure in the unbound state that efficiently quenches the fluorescence signal at all temperatures.

The result is a very low background fluorescence and consequently higher signal-to-noise ratio compared to standard hydrolysis probes.

HydrolEasy® probes are highly suitable for multiplexed real-time PCR applications where multiple probes with the same fluorophore are used.

  • Increased signal-to-noise ratio
  • Improved target affinity and specificity
  • Unbound probe is quenched at all temperatures
  • Available with a wide range of fluorophores and quenchers
  • Ideal for same-color multiplexed real-time PCR applications

1 TaqMan® is owned by Roche Diagnostics, Inc.

Product Details

Reference Numbers                                                459-478

Product Name                                                          HydrolEasy® probe 14-35 mer

Flourophores                                                            More than 35 fluorophores including PentGreen/Yellow/Orange/Red, CAL Flour, Quasar, ATTO and CYanine – see catalogue for details

Purification                                                                HPLC

Yield                                                                           10/50 nmol or higher delivered

Format                                                                        Lyophilized                                                               

Verification                                                                Melt analysis using complementary target


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Related Files
File name
Customised Oligos and PCR Assays Catalogue V.2.5

Naturally Quenched

By adding hydrophobic IPN molecules close to the ends of the dual-labelled HydrolEasy® probe, the fluorophore and quencher are automatically brought into close proximity due to interactions between IPN molecules. This leads to quenching of the unbound probe at all temperatures.

The addition of IPN molecules to HydrolEasy® probes is automatically done by PentaBase when receiving an order and is included in the price. The number of added IPN molecules depends on the desired melting temperature of the probe. The addition of IPN molecules typically adds 4-8°C to the melting temperature.

Increased Signal-to-noise Ratio

The lower background fluorescence of HydrolEasy® probes compared to standard TaqMan® probes leads to higher signal-to-noise (S/N) ratio of normalized flourescence.

This is particularly useful in assays that target low abundant and/or less accessible genetic regions by providing relative fluorescence levels above minmum requirements that would otherwise be difficult to reach with standard hydrolysis probes.

Inceased Target Affinity

HydrolEasy® probes have increased target affinity compared to standard TaqMan® probes. The graphs show melt curves (top) and melt peaks (bottom) of a TaqMan® probe and a corresponding HydrolEasy® probe (bold lines) with identical sequence.

The melting temperature (+ target) of the HydrolEasy® probe is significantly higher than the corresponding TaqMan® probe.

The background fluorescence (- target) of the HydrolEasy® probe is also significantly lower than the corresponding TaqMan® probe resulting in a considerably higher melting peak height.

These features of HydrolEasy® probes can be used to design PCR assays with higher sensitivity and specificity compared to assays based on standard TaqMan® probes.

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