DescriptionXRN-1 is highly processive 5´→3´ exoribonuclease, requiring 5´ monophosphate. It also acts on 5´ monophosphate ssDNA with greatly reduced efficiency. XRN-1 is not efficient in removing RNAs with recessed 5´P, like tRNA. It does not cleave dsDNA, ssDNA and RNA which contain di, triphosphate, OH and capped 5´ ends.
Product SourcePurified from E. coli carrying a plasmid overexpressing the yeast XRN-1 gene (1).
The following reagents are supplied with this product:
|Store at (°C)||Concentration|
Properties and Usage
Unit DefinitionOne unit is defined as the amount of enzyme that digests 1 µg of phosphorylated yeast RNA in 60 minutes at 37°C.
1X NEBuffer 3
Incubate at 37°C
1X NEBuffer 3:
100 mM NaCl
50 mM Tris-HCl
10 mM MgCl2
1 mM DTT
pH 7.9 @ 25°C
20 mM Tris-HCl
500 mM NaCl
2 mM DTT
0.1 mM EDTA
0.1% Triton® X-100
Heat Inactivation70°C for 10 min
Quality Control AssaysThe following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page.
- Endonuclease Activity (Circular Single Stranded DNA):
The product is tested in a reaction containing circular single stranded DNA. After incubation the percent cleaved is determined by agarose gel electrophoresis.
- Endonuclease Activity (Nicking):
The product is tested in a reaction containing a supercoiled DNA substrate. After incubation for 4 hours the percent converted to the nicked form is determined by agarose gel electrophoresis.
- RNase Activity (2 Hour Digestion):
The product is tested in a reaction containing a RNA substrate. After incubation for 2 hours there is no detectable degradation of the RNA substrate as determined by gel electrophoresis.
Material Safety Datasheets
- We recommend adding Murine RNase Inhibitor (NEB #M0314) in your reaction at 1 unit/µl to prevent non-specific RNA degradation.
- Stevens, A (1980). Biol. Chem. 255, 3080-3085.