Recombinant RNase R (E. coli) is a magnesium-dependent 3′→5′ exoribonuclease that can digest all linear RNAs but does not digest lariat or circular RNA structures. Most cellular RNAs will be digested completely by RNase R, with the exception of tRNAs, 5S RNA and intron lariats. RNase R is used for enrichment of circRNAs in biological samples, identification of intronic lariat sequences, identification of exonic circRNAs, studying alternative splicing.
Buffer: 50mM Tris-HCl, 200mM NaCl, 1mM DTT, 0.1mM EDTA, 0.1% Triton X-100, 50% glycerol, pH=7.5.
Storage: -20℃
Instructions
Prepare the following reaction system in a sterile microcentrifuge tube:
|
Component |
Volume |
|
1×RNase R Reaction Buffer |
20 μL |
|
RNA sample |
1 μg |
|
RNase R (20 U/μL) |
2-4 U |
|
RNase-free ddH2O |
Up to 20 μL |
Notes
1. The activity of RNase R requires 0.1-1.0 mM Mg2+.
2. With the increase of RNA, the reaction time can be extended and the enzyme amount can be increased appropriately.
3. The EDTA content in RNA samples may affect the activity of RNase R.
4. Incubate at 70 ℃ for 10 minutes can inactivate the enzyme.
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