RNase R (20 U/μL) -14606ES

Ribonuclease R (RNase R) from 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.

Specificntions

Source	A recombinant E. coli strain carrying the RNAse R gene from E. coli
Optimum Temperature	37℃
Storage Buffer	50 mM Tris-HCl,100 mM NaCl, 0.1 mM EDTA, 0.1% Triton® X-100, 1 mM DTT, 50% glycerol，pH7.5 at 25℃
Unit Definition	One unit converts 1 μg of poly-r(A) into acid-soluble nucleotides in 10 minutes at 37°C in 20 mM Tris-HCl (pH 8.0), 100 mM KCl and 0.1 mM MgCl2.

Components

Components No.	Name	14606ES72	14606ES86	14606ES92
14606-A	RNase R (20 U/μL)	12.5 μL	125 μL	500 μL
14606-B	10×RNase R Reaction Buffer	250 μL	3 × 1 mL	10 mL

Storage

This product should be stored at -25~-15℃ for 2 years.

Instructions

Prepare the following reaction system in a sterile microcentrifuge tube

component	Volume
10×RNase R Reaction Buffer	2 μL
RNA sample	1 μg
RNase R (20 U/μL)	2-4 U
RNase-free ddH2O	Up to 20 μL

Incubate at 37 ℃ for 10 - 30 minutes.

Figures

Protein purity (SDS-PAGE) ≥95% 

Figure 1: Protein purity detection: Different volumes (1 μL, 2 μL, 3 μL) of the product (Cat#14606ES) show high purity ≥95%.
 

No residual exonuclease

 

Figure 2: 20 U of RNASE R was added to 0.5 μg of λDNA-Hind III Digest, incubating at 37 ℃ for 4 hours. The result shows that no residual exonuclease was detected from the RNASE R (CAT#14606ES) products.

Ability of digestion linear RNA and CircRNA

Figure 3: RNASE R (CAT#14606ES) can digest linear RNA molecules

Figure 4: CircRNA can tolerate the treatment of RNASE R (CAT#14606ES).

 

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.

 

 

Documents:

Manuals

14606ES-RNase R (20 UμL)-HB230531.pdf