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A major application for Ribonuclease A (RNase A) is the removal of RNA from preparations of plasmid DNA. In this application, the presence of DNase activity as an impurity is a concern. The boiling-water bath method used to eliminate contaminating DNase activity has proven unreliable. For this reason, developed a proprietary chromatographic preparation method for elimination of DNase activity.
RNase A is an endoribonuclease that attacks at the 3¢ phosphate of a pyrimidine nucleotide. The sequence of pG-pG-pC-pA-pG will be cleaved to give pG-pG-pCp and A-pG. The highest activity is exhibited with single stranded RNA. RNase A is a single chain polypeptide containing 4 disulfide bridges. In contrast to RNase B, it is not a glycoprotein. RNase A can be inhibited by alkylation of His12 or His119 , which are present in the active site of the enzyme. Activators of RNase.
Storage/Stability
This product remains active for at least 3 years when stored properly at –20 °C. RNase A is a very stable enzyme and solutions have been reported to withstand temperatures up to 100 °C. At 100 °C, an RNase A solution is most stable between pH 2.0 and 4.5.
Procedure
A major application for RNase A is the removal of RNA from preparations of plasmid DNA. For this application, DNase free RNase A is used at a final concentration of 0.2 mg/ml.
Mr. Rakesh Singh (BR Biochem Life Sciences Pvt. Ltd.)
1/10, First Floor, Tilak Nagar, Delhi - 110018, India
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