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Enzymes for Molecular Biology

Enzymes for Molecular Biology: Enzymes for Molecular Biology

RNase R

RNase R

CellScript

Catalogue No.DescriptionPack SizePriceQty
RRT250325RNase R25 µg £106.00 Quantity Add to Order

Description

Ribonuclease R (RNase R) from E. coli, is a 3’→5’ exoribonuclease that can digest linear RNA and Y-structure RNA, but does not digest circular RNA (circRNA), lariat intron RNA, blunt-ended double-stranded RNA (dsRNA), dsRNA with 3’-overhangs <7 nt in length or single-stranded DNA.1,2 Highly structured linear cellular RNAs such as prokaryotic 23S and 16S ribosomal RNAs (rRNAs), small nuclear RNAs (snRNAs), transfer RNAs (tRNAs) and histone mRNAs are also not efficiently degraded by RNase R.3 This functionality makes RNase R ideal for isolating circular RNA structures from total cellular RNA preparations for use in intron and splicing studies and natural cellular circRNA identification. Additionally, RNase R is used as a crucial component in in vitro circRNA preparation cleanup and enrichment3-5 with both RNA Ligase-based6 or self-splicing permutated intron–exon sequence (PIE method)-based5,7 methodologies.

 

Product Performance:

Figure 1. A PIE-method RNA circularization prep was assessed on a denaturing gel before (UT) and after (T) RNase R treatment. The untreated sample lane shows different bands corresponding to linear RNA subproducts from the PIE-method. Following RNase R treatment, the linear RNA are digested, leaving only the circular RNA (red arrow) intact.

Fig 1_PIE method RNA circularization

 

 

Figure 2. 5 µg of linear RNA was circularized with RNA Ligase to produce circular RNA (lane 2). The RNA ligation reaction product was treated with 1 µg of RNase R at 37°C for 30 minutes to enrich for circular RNAs (lane 3).

Fig 2_RNA Ligase method RNA circularization

Materials Supplied:

Important Store at -20°C in a freezer without a defrost cycle. Do not store at –70°C.

 

 

Terms & Conditions: The Product listed is currently offered by CELLSCRIPT™ for research use only under the defined Terms and Conditions.

 

FAQs: For answers to Frequently Asked Questions about our products, visit the FAQ library.

Troubleshooting Guide:

 

 

Symptom Solution
All linear RNA is not degraded Increase the time of incubation of the reaction.
Increase the amount of RNase R used in the reaction.
Decrease the amount of input RNA in the reaction.
RNA contains stable secondary structure whether internally or at the 3' end.

• Add a short poly(A) tail onto the RNA to provide a structureless 3' end starting point for RNase R binding.3
• Note: some highly stable secondary structures are resistant to RNase R digestion.

 

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

Protocols

These are both links to the manufacturer's page

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

References

1. Suzuki, H. et al., (2006) Nucl. Acids Res. 34 (8) e63.

2. Vincent, H.A. and Deutscher, M.P., (2006) J. Biol. Chem. 281 29769.

3. Xiao, M.S. and Wilusz, J.E., (2019) Nucl. Acids Res. 47 8755.

4. Wesselhoeft, R.A. et al., (2018) Nat Commun. 9 2629.

5. Qui, Z. et al., (2022) BioRxiv. doi: 10.1101/2022.06.20.496777.

6. Beaudry, D. and Perreault, J.P., (1995) Nucl. Acids Res. 23 3064.

7. Puttaraju, M. and Been, M.D., (1996) J. Biol. Chem. 271 26081

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

Applications & Benefits

  • Digests linear RNA and Y-structure RNA, leaving circRNA, lariat intron RNA, and dsRNA intact.
  • Ideal for isolating circRNA from total RNA preparations.
  • 1 µg of RNase R digests 5 µg of linear RNA in 30 minutes.

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200