94 kD
50mM Tris-HCl, 200mM NaCl, 1mM DTT, 0.1mM EDTA, 50% (v/v) Glycerol, 0.1% (w/v) Triton X-100 (pH7.5 @25℃)
Store at -25 ~ -15℃ for 1 years
[1] Cheng, Z.-F. Purification and Characterization of the Escherichia coli Exoribonuclease RNase R COMPARISON WITH RNase II[J]. Journal of Biological Chemistry, 2002, 277(24):21624.
[2] Cheng Z F, Deutscher M P. An Important Role for RNase R in mRNA Decay[J].Molecular Cell, 2005, 17(2):313-318.
Ribonuclease R (RNase R) is a Mg2+ dependent 3'→5' exonuclease derived from Escherichia coli. RNase R can digest all linear RNA. However, it cannot digest ring-shaped RNA, lasso structure or double-stranded RNA molecules with 3 'end protruding ends less than 7 nucleotides, tRNA and 5SRNA with complex secondary structure. RNase R is commonly used in gene expression and variable shear studies and can digest linear RNA to enrich circular RNA or lasso structured RNA. This product does not contain DNase, other RNA endonuclease and exonuclease activities.
Storage Solution: 20U/μl RNase R, 50mM Tris-HCl, 200mM NaCl, 1mM DTT, 0.1mM EDTA, 50% (v/v) Glycerol, 0.1% (w/v) Triton X-100 (pH7.5 @25℃) 10*Reaction Buffer: 200mM Tris-HCl, 1M KCl, 1mM MgCl2 (pH8.0 @25℃)
1、Set up the following reaction in a microcentrifuge tube on ice. Add the following components in sequence.
Components | Volume 20μl |
10* Reaction Buffer | 2 μl |
RNA sample | 1 μg |
RNase R (20 U/μL) | 2-4 U |
Up to 20 μL |
2、Reaction at 37℃ for 10 min-30 min.
1. The activity of RNase R requires 0.1-1.0 mM Mg2+;
2. With the increase of substrate RNA, digestion time and enzyme amount can be appropriately prolonged
In the 20µl reaction system, the mixture of linear and circular RNA was used as substrate, nuclease-free water was added to the control group, RNase R was added to the experimental group, the reaction was performed at 37℃ for 30min. The reaction was analyzed by 1% agarose gel electrophoresis.
M: DNA marker
Lane 1 negative control
Lane 2 Experimental result
1μg (R: reducing condition, N: non-reducing condition).
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