RiboCleave™ 1
Sequence-specific RNA restriction enzyme
RiboCleave™ 1
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RiboCleave™ 1 is a sequence-specific RNA restriction enzyme that recognizes and cleaves the GAA↓AU sequence in single-stranded RNA. Designed for RNA analytics applications, it enables rapid generation of defined analytical fragments for mRNA cap-efficiency assessment. A single digestion step replaces complex multi-step RNase H workflows, reducing sample preparation time from approximately 6 hours to less than 30 minutes. As a sample-preparation step, RiboCleave™ 1 is method-independent by design: the same digest can be analyzed by urea-PAGE, Bioanalyzer or LC-MS.
Key Benefits
- Reduces cap-analysis sample preparation from approximately 6 hours to less than 30 minutes.
- Eliminates guide oligo design and optimization requirements.
- Method-independent by design: one digest for urea-PAGE, Bioanalyzer and LC-MS readouts.
- Enables in-house cap analysis on standard gel equipment.
Key Features
- Sequence-specific cleavage of GAA↓AU motifs in single-stranded RNA.
- Single-enzyme, one-incubation workflow.
- Compatible with natural and modified RNA substrates, including N1-methylpseudouridine and pseudouridine.
- Supplied with dedicated Reaction Buffers for analytical flexibility on gel and/or HPLC/LC-MS readouts
Figures
The activity was measured over 10 min of reaction at 37°C in a 25 mM Tris-HCl buffer, pH 8.0, 0.1 µM Quench1 substrate and 1x DGB buffer (final 1 mM DTT, 2 mM Glycerol and 0.2 mg/ml BSA) and with varying concentrations of NaCl.
Average of 2 experiments.
The activity was measured over 10 min of reaction at 37°C in a 25 mM Tris-HCl pH 8.0, 25 mM NaCl, 0.1 µM Quench1 substrate, 1x DGB buffer (final 1 mM DTT, 2 mM Glycerol and 0.2 mg/ml BSA) and varying MgCl2 concentrations.
Average of 2 experiments.
The activity was measured over 10 min of reaction at 37°C in a 25 mM Tris-HCl buffer, 25 mM NaCl, 0.1 µM Quench1 substrate and 1x DGB buffer (final 1 mM DTT, 2 mM Glycerol and 0.2 mg/ml BSA) and varying pH.
Average of 2 experiments.
The activity was measured after 10 min of reaction in a 25 mM Tris-HCl pH 8.0, 25 mM NaCl, 0.1 µM Quench1 substrate and 1x DGB buffer (final 1 mM DTT, 2 mM Glycerol and 0.2 mg/ml BSA) and varying temperature.
Average of 2 experiments.
The activity was measured over 10 min of reaction at 37°C in a 25 mM Tris-HCl buffer, pH 8.0, 0.1 µM Quench1 substrate and 1x DGB buffer (final 1 mM DTT, 2 mM Glycerol and 0.2 mg/ml BSA) and with varying concentrations of ammonium acetate.
Average of 2 experiments.
The activity was measured over 10 min of reaction at 37°C in a 25 mM Tris-HCl pH 8.0, 25 mM NaCl, 0.1 µM Quench1 substrate, 1x DGB buffer (final 1 mM DTT, 2 mM Glycerol and 0.2 mg/ml BSA) and varying MnCl2 concentrations.
Average of 2 experiments.
10 pmol of RNA oligo bearing known star site in 10 µL water were mixed with 10 µl of 2x Reaction Buffer A composed of 25 mM Tris-HCl pH 8.5 @ 25°C, 25 mM NaCl,and 1 µl Ribocleave 1 at 1 U/µl. Reaction was incubated for 10 min at 37°C.
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