New England Biolabs, M0338S, XRN-1

XRN-1 is highly processive 5´→3´ exoribonuclease, requiring 5´ monophosphate. It also acts on 5´ monophosphate ssDNA with greatly reduced efficiency. Related Categories RNases,, Epitranscriptome Analysis,, RNA Modification Specification 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. Reaction Conditions 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) Storage Buffer 20 mM Tris-HCl 500 mM NaCl 2 mM DTT 0.1 mM EDTA 50% Glycerol 0.1% (w/v) Triton® X-100 FAQ Q: Can XRN-1 digest ssDNA containing 5’ monophosphates? A: Yes. However, it will cleave ssDNA at a lower efficiency than RNA containing 5’ monophosphate. Q: Is XRN-1 a tagged protein? A: No, XRN-1 is not a tagged protein. Q: Is XRN-1 identified with other names in the literature? A: Yes. XRN1 has been referred to as KEM1, SEP1, DST2, RAR5 SKI1 and DST2 in the literature. Q: Is XRN-1 salt tolerant? A: No significant activity loss is observed at 400 mM salt. Q: What is the molecular weight of XRN-1? A: XRN-1 has a molecular weight of 175 kDa. Q: Is it possible to digest RNase R (NEB #M0100)-resistant linear RNAs? A: Yes, we have tested two methods to digest RNase R-resistant linear RNAs: We recommend adding 1 unit of RNase R, 1 unit of XRN-1 (NEB #M0338), and 5 units of RppH (NEB #M0356) to 1 µg of RNA sample in 1X RNase R Reaction Buffer. The reaction should be incubated for 15 or 30 minutes at 37°C for efficient linear RNA removal. Pre-treat RNA samples with E. coli Poly(A) Polymerase (NEB #M0276) to lengthen the 3´ ends of the RNAs, which can improve RNase R binding and activity. The RNA needs to be cleaned up before proceeding with the RNase R reaction.