Rnase A

Ribonuclease A (RNaseA) is a single-stranded polypeptide containing 4 disulfide bonds with a molecular weight of about 13.7 kDa. RNase A is an endoribonuclease that specifically degrades single-stranded RNA at C and U residues. Specifically, the cleavage recognizes the phosphodiester bond formed by the 5′-ribose of a nucleotide and the phosphate group on the 3′-ribose of the adjacent pyrimidine nucleotide, so that the 2,3′-Cyclic phosphates are hydrolyzed to the corresponding 3′nucleoside  phosphates(eg,pG-pG-pC-pA-pG is cleaved by RNase A to generate pG-pG-pCp and A-PG). RNase  A is the most active in cleaving single-stranded RNA.Recommended working concentration is 1-100μG/mL,compatible with various reaction systems.Low salt concentration (0-100 mM NaCl)can be used to cut single-stranded RNA,double-stranded RNA,and RNA chains formed by RNA-DNA hybridization.However, at high salt concentration (≥0.3 M), RNase A only specifically cleaves single-stranded RNA.

Storage Conditions

The product can be stored at -25～-15℃, valid for 2 years

Instructions

This is one of the common methods for preparing RNase A storage solution.It can also be prepared by other methods according to the traditional methods in the laboratory or reference literature (such as directly dissolving in 10 mM Tris-HCl, pH 7.5 or Tris-NaCl solution) 

1.Use 10 mM sodium acetate (pH5.2)to prepare 10 mg/mL of RNase A storage solution

2.Heating at 100℃ for 15 min

3.Cool to room temperature,add 1/10 volume of 1 M Tris-HCl (pH 7.4),adjust its pH to 7.4(for example,add 500 ml of 10g/ml RNase storage solution 1M Tris-HCL, PH7.4）

4.Sub-packaged at-20℃ for frozen storage,which can be stable for up to 2 years

[Notes]: When boiling RNaseA solution under neutral conditions,RNase precipitation will

form;Boil it at a lower pH, and if there is precipitation,it can be observed,which may be

caused by the presence of protein impurities.If sediment is found after boiling,impurities can

be removed by high-speed centrifugation (13000 rpm), and then sub- packed for freezing storage

Product information

Notes

For your safety and health, please wear lab coats and disposable gloves for operation.