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M0100 RNase R

Catalog # Concentration Size List Price Quantity Your Price
M0100S 400 units $153.00
$137.70
Catalog # Size List Price Your Price
M0100S 400 units $153.00
$137.70
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca
  • Highly processive 3´ to 5´ exoribonuclease
  • 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

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

Mechanism of RNase R, showing digestion of linear RNA and tail of lariat RNA, but no access to circular RNA

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

Gels showing optimal performance of RNase R for linear RNA, circular RNA and both

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.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  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
An exception occurred during the operation, making the result invalid. Check InnerException for exception details.

Companion Products
References
  • 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
Quality Control Assay
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Specifications
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
Certificate of Analysis
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Safety Data Sheets
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