Digests nearly all linear RNAs with an accessible 3´ end, including rRNA and mRNA
Enriches for circular RNAs and lariat RNAs from total RNA preps
Requires magnesium for activity
Inactivated with the addition of excess EDTA
Product Information
Ribonuclease R (RNase R) is a processive 3´ to 5´ exoribonuclease (1,2). RNase R requires a 3´ single-stranded RNA substrate that is approximately 10 nucleotides or longer, such as a poly (A) tail, for proper binding (3,4,5). RNase R is unique from other exoribonucleases for its ability to degrade highly structured RNAs without the need for an additional helicase (2,6,7). Circular RNAs and lariat RNAs are enriched after RNase R digestion as these closed RNA structures are resistant to exoribonucleases (8).
Figure 1. RNase R digests linear RNA with an accessible 3´ end
RNase R digests linear RNA with an accessible 3´ end and can be used to enrich for circular and lariat RNA. RNase R can be used to digest linear RNAs. Circular RNAs are closed RNA molecules that are resistant to RNase R digestion. Lariat RNAs have a looped structure with a single-stranded RNA region at the 3´ end. RNase R can digest the linear RNA at the 3´ end, but stops at the branch point of the lariat, preserving the looped RNA.
Figure 2. RNase R efficiently degrades linear RNA and leaves circular RNA intact
NEB® RNase R efficiently degrades linear RNA and leaves circular RNA intact. Adding 1 unit of NEB RNase R per 1 µg of RNA sample containing only linear RNA, only circular RNA, or both linear and circular RNA results in circular RNA maintenance and linear RNA digestion. This reaction ran for 15 or 30 minutes at 37°C in 1X RNase R Reaction Buffer.
Product Source
An E. coli strain that carries the cloned RNase R gene (rnr) from Escherichia coli with an N-terminal 6xHis-tag.
This product is related to the following categories:
The following reagents are supplied with this product:
NEB #
Component Name
Component #
Stored at (°C)
Amount
Concentration
M0100S
-20
RNase R
M0100SVIAL
-20
1 x 0.02 ml
20,000 units/ml
RNase R Reaction Buffer
B0100SVIAL
-20
2 x 1.25 ml
10 X
Properties & Usage
Unit Definition
One unit is defined as the amount of enzyme required to convert 75 pmoles of 20-nucleotide single-stranded RNA sequence downstream of a 38-nucleotide DNA hairpin into acid soluble ribonucleotides in a total reaction volume of 20 µl in 15 minutes at 25°C.
Reaction Conditions
1X RNase R Reaction Buffer
Incubate at 37°C
1X RNase R Reaction Buffer
20 mM Tris-HCl
100 mM KCl
0.1 mM MgCl2
Storage Buffer
300 mM NaCl
10 mM Tris-HCl
0.1 mM EDTA
1 mM DTT
surfactant
50% Glycerol
pH 7.4 @ 25°C
Kasai T, et al. (1977). Exoribonucleases in wild type Escherichia coli and RNase II-deficient mutants. J Biol Chem. Dec 25; 252 (24), 8950-6. PubMedID: 336626
Cheng ZF and Deutscher MP. (2002). Purification and characterization of the Escherichia coli exoribonuclease RNase R. J Biol Chem. Jun 14; 277 (24), 21624-9. PubMedID: 11948193
Cheng ZF and Deutscher MP. (2005). An important role for RNase R in mRNA decay. Mol Cell. Jan 21; 17(2), 313-8. PubMedID: 15664199
Vincent HA and Deutscher MP. (2006). Substrate recognition and catalysis by the exoribonuclease RNase R. J Biol Chem. Oct 6;281(40), 29769-75. PubMedID: 16893880
Xiao MS and Wilusz JE. (2019). An improved method for circular RNA purification using RNase R that efficiently removes linear RNAs containing G-quadruplexes or structured 3' ends. Nucleic Acids Res. Sep 19; 47 (16), 8755-69. PubMedID: 31269210
Hossain ST, et al. (2016). How RNase R Degrades Structured RNA: ROLE OF THE HELICASE ACTIVITY AND THE S1 DOMAIN. J Biol Chem. Apr 8; 291 (15), 7877-87. PubMedID: 26872969
Awano N, et al. (2010). Escherichia coli RNase R has dual activities, helicase and RNase. J. Bacteriol. Mar; 192 (5), 1344-52. PubMedID: 20023028
Suzuki H, et al. (2006). Characterization of RNase R-digested cellular RNA source that consists of lariat and circular RNAs from pre-mRNA splicing. Nucleic Acids Res. May 8; 34 (8), e63. PubMedID: 16682442
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Specifications & Change Notifications
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