Ribonuclease R (RNase R)
Cat. No.: SB-G3462-500U
Size: 500U
Description
Ribonuclease R, referred to as RNase R, is a 3'-5' exoribonuclease derived from Escherichia coli, which is recombinantly expressed in Escherichia coli, and gradually cuts RNA into dinucleotides and trinucleotides from the 3'-5' direction . It can degrade almost all linear RNA molecules and Y-structure RNA, but it is not easy to degrade circular RNA (circRNA), lariat structure (lariat RNA) and double-stranded RNA molecules with 3' protruding ends of less than 7 nucleotides. Based on its characteristics, it is mainly used for gene expression and variable splicing research, enriching circular RNA or lariat RNA.
- Source : derived from Escherichia coli, recombinantly expressed by Escherichia coli.
- Definition of enzyme activity : under standard reaction conditions at 37°C, the amount of enzyme required to convert 1 μg poly(A) into acid-soluble nucleotides within 10 minutes is defined as one enzyme activity unit.
- Purity and concentration : SDS-PAGE detection purity ≥ 95%; 20 U/μL.
- Enzyme storage buffer : 50 mM Tris-HCl, 100 mM NaCl, 0.1 mM EDTA, 1 mM DTT, 0.1% Triton X-100, 50% Glycerol, pH7.5.
- 10 × Reaction Buffer : 200 mM Tris-HCl, 1 M KCl, 1 mM MgCl2 , pH 8.0.
Storage and Handling Conditions
Transport in wet ice packs; store at -20°C, valid for 12 months.
Composition
| Component Number |
Component |
G3462-500U |
| G3462-1 |
Ribonuclease R |
25 μL |
| G3462-2 |
10 × Reaction Buffers |
500 μL |
| Product Manual |
1 copy |
Steps
1. Configure the reaction system according to the following table:
| Component |
Volume |
| RNA sample |
＜5μg |
| 10 × Reaction Buffers |
2 μL |
| RNase R |
0.5 μL (1-3 U/μg RNA) |
| Nuclease Free Water |
To 20 μL |
Note: The amount of RNase R can be adjusted according to the actual situation.
2. After the reaction system is fully mixed, incubate at 37°C for 10-30 min. (Note: The reaction time is not easy to exceed 1 h)
3. After the reaction is complete, incubate at 70°C for 10 minutes to terminate the reaction. The product can be purified or directly subjected to subsequent RT-PCR experiments.
Precautions
- All enzymes should be stored in an ice box and stored at -20°C immediately after use.
- For your safety and health, please wear a lab coat and disposable gloves for operation.
