A processive 5´ to 3´ riboexonuclease that specifically digests RNA with 5´-monophosphate ends
- Specific: Processively digests RNA with 5´-monophosphate ends but not RNAs with 5´-triphosphate, 5´-cap or 5´-hydroxyl groups starting from the 5´ end
- Flexible: Use this enzyme to help characterize 5´-ends of RNA and to enrich for mRNA in prokaryotic or eukaryotic total RNA preparations
Terminator™ 5´-Phosphate-Dependent Exonuclease* is a processive 5´ to ;3´ exonuclease that digests RNA that has a 5´-monophosphate end. It does not digest RNA that has a 5´-triphosphate, 5´-cap or 5´-hydroxyl group. Terminator Exonuclease is not inhibited by proteinaceous RNase inhibitors, such as RiboGuard™ RNase Inhibitor or placental ribonuclease inhibitors.
Applications
- Characterize the 5´ termini of RNA transcripts.
- Enrich for mRNA in prokaryotic or eukaryotic total RNA preparations (Fig. 1).
Note: Terminator™ 5´-Phosphate-Dependent Exonuclease has not been optimized for RNA-Seq applications.
Unit Definition: One unit of Terminator™ Exonuclease digests 1 µg of rRNA substrate to acid-soluble nucleotides in 60 minutes at 30°C under standard assay conditions.
Storage Buffer: 50% glycerol containing 0.05 M Tris-HCl (pH 7.5), 100 mM NaCl, 0.1 mM EDTA, 1 mM DTT, and 0.1% Triton™ X-100.
Terminator™ 5´-Phosphate-Dependent Exonuclease* is a processive 5´ to ;3´ exonuclease that digests RNA that has a 5´-monophosphate end. It does not digest RNA that has a 5´-triphosphate, 5´-cap or 5´-hydroxyl group. Terminator Exonuclease is not inhibited by proteinaceous RNase inhibitors, such as RiboGuard™ RNase Inhibitor or placental ribonuclease inhibitors.
Applications
- Characterize the 5´ termini of RNA transcripts.
- Enrich for mRNA in prokaryotic or eukaryotic total RNA preparations (Fig. 1).
Note: Terminator™ 5´-Phosphate-Dependent Exonuclease has not been optimized for RNA-Seq applications.
Unit Definition: One unit of Terminator™ Exonuclease digests 1 µg of rRNA substrate to acid-soluble nucleotides in 60 minutes at 30°C under standard assay conditions.
Storage Buffer: 50% glycerol containing 0.05 M Tris-HCl (pH 7.5), 100 mM NaCl, 0.1 mM EDTA, 1 mM DTT, and 0.1% Triton™ X-100.
Figure 1. A 1-hour Terminator™ Exonuclease reaction digests the large rRNAs in a eukaryotic or prokaryotic total RNA sample, producing an enriched mRNA preparation. |
Figure 2. Normal rat kidney (NRK) total RNA before (A) and after (B) Terminator™ Exonuclease treatment. The Terminator Exonuclease-treated RNA was concentrated 10-fold. | Figure 3. Denaturing agarose gel analysis of E. coli total RNA before (-) and after (+) Terminator™ Exonuclease digestion. The Terminator Exonuclease-treated RNA was concentrated 10-fold. |
Figure 4.