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XRN-1 |
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Prices are in US dollars and valid only for US orders.
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Description:
XRN-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.
Source:
Purified from E. coli carrying a plasmid overexpressing the yeast XRN-1 gene (1).
Applications:- Removal of RNA containing 5´ monophosphate from a RNA mixture.
Reagents Supplied:
NEBuffer 3 (10X)
Enzyme Properties
Heat Inactivation:
70°C for 10 minutes
Reaction & Storage Conditions
Reaction Conditions:
1X NEBuffer 3
Incubate at
37°C.
1X NEBuffer 3:
50 mM Tris-HCl
100 mM NaCl
10 mM MgCl2
1 mM Dithiothreitol
pH 7.9 @ 25°C
Unit Definition:
One unit is defined as the amount of enzyme that digests 1 µg of phosphorylated yeast RNA in 60 minutes at 37°C.
Concentration:
1,000 units/ml
Storage Conditions:
20 mM Tris-HCl
500 mM NaCl
2 mM Dithiothreitol
0.1 mM EDTA
50% Glycerol
0.1% Triton X-100
Storage Temperature:
-20°C
Notes
Usage notes:- We recommend adding Murine RNase Inhibitor (NEB #M0314) in your reaction at 1 unit/µl to prevent non-specific RNA degradation.
FAQs
- What is the molecular weight of XRN-1?
- Is XRN-1 a tagged protein?
- Can XRN-1 digest ssDNA containing 5’ monophosphates?
- Can XRN-1 digest RNA that contains a di or triphosphate at 5’end?
- Is XRN-1 identified with other names in the literature?
Quality Control for Current Lot
Quality control values for a specific lot can be found on the datacard which accompanies each product.
RNase Assay::
Incubation of 2 units of XRN-1 with 40 ng of RNA transcript in 10 µl for 2 hours at 37°C resulted in no detectable degradation of RNA as determined by denaturing PAGE analysis.
Endonuclease Activity: dsDNA::
Incubation of 2 units of XRN-1 with 300 ng of supercoiled plasmid in 10 µl for 4 hours at 37°C produced less than 10% nicked or linear molecules as determined by agarose gel electrophoresis.
Endonuclease Activity: ssDNA::
Incubation of 2 units of XRN-1 with 250 ng of M13 SSDNA in 10 µl for 4 hours at 37°C resulted in no detectable degradation as determined by agarose gel electrophoresis.
References
- Stevens, A. (1980) Biol. Chem, 255, 3080-3085.
Reagents Sold Separately
NEBuffer 3
Companion Products
Murine RNase Inhibitor
RNase Inhibitor
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