Enzymes for Molecular Biology: Enzymes for Molecular Biology

Terminator™ 5"-Phosphate-Dependent Exonuclease* (Terminator Exonuclease) is a 5"-to-3", processive exonuclease that degrades RNAs that have a 5"-monophosphate - such as prokaryotic 16S and 23S rRNA and eukaryotic 18S and 28S rRNA. RNAs with a 5"-hydroxyl group, RNAs with a 5"-cap structure (such as that found on most eukaryotic mRNAs) and RNAs with a 5-triphosphate (such as that found on the 5"-end of primary prokaryotic RNA transcripts) are resistant to degradation by Terminator Exonuclease. Thus, Terminator Exonuclease is ideal for producing an mRNA-enriched sample from a total RNA preparation by selectively removing the large rRNAs.

Terminator Exonuclease will also digest DNA with a 5"-monophosphate. Terminator Exonuclease is not inhibited by proteinaceous RNase inhibitors, such as RNasin® (Promega Corp) or other placental ribonuclease inhibitors, or by Prime® Inhibitor (Eppendorf).

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 mMEDTA, 1 mM DTT, and 0.1% Triton™ X-100.

Important information

Occassionally Terminator Exonuclease displays RNAse activity when using Buffer A (in a bacterial rRNA removal). We would recommend the Ribozero kits if using bacterial species, particularly for sequencing purposes.

If you cannot find the answer to your problem below then please contact us or telephone 01954 210 200

Protocols for: Terminator™ 5"-Phosphate-Dependent Exonuclease

Protocol

(catalogue number TER51020)

Please note: all protocols off site are the responsibility of the products supplier

If you cannot find the answer to your problem below then please contact us or telephone 01954 210 200

Important information

Occassionally Terminator Exonuclease displays RNAse activity when using Buffer A (in a bacterial rRNA removal). We would recommend the Ribozero kits if using bacterial species, particularly for RNA sequencing purposes.

If you cannot find the answer to your problem below then please contact us or telephone 01954 210 200

Applications

• Characterize the 5´ termini of RNA transcripts.
• Enrich for mRNA in prokaryotic or eukaryotic total RNA preparations

If you cannot find the answer to your problem below then please contact us or telephone 01954 210 200