The RNase A is free of DNase activity. It is not necessary to heat it before use.

The RNase A, DNase and pretease-free is an endoribonuclease that specifically degrades single-stranded RNA at C and U residues. It cleaves the phosphodiester bond between the 5¡¯-ribose of a nucleotide and the phosphate group attached to the 3¡¯-ribose of and adjacent pyrimidine nucleotide. The resulting 2¡¯, 3¡¯-cyclic phosphate is hydrolyzed to the corresponding 3¡¯-nucleoside phosphate.

*  Plasmid and genomic DNA preparation

*  Removal of RNA from recombinant protein preparations.

*  Ribonuclease protection assays

*  Mapping single-base mutation in DNA or RNA

 

The absence of endodeoxyribonucleases, exodeoxyribonucleases and proteases confirmed by appropriate quality tests. Functionally tested for RNA digestion in a plasmid DNA purification procedure.

 

Bovine pancreas.

 

13.7 kDa monomer

 

One unit of the enzyme causes an increase in absorbance of 1.0 at 260 nm when yeast RNA is hydrolyzed at 37¡É and pH 5.0. Fifty units are approximately equivalent to 1 Kunitz unit.

 

>5000 u/mg protein (>100 Kunitz/mg protein)

 

 

The enzyme is supplied in: 50 mM Tris-HCl(pH 7.4) and 50% (v/v) glycerol.

*  Inhibitors: the most potent inhibitor is 1~50 kDa protein from cytosol of mammalian cells, e.g., Mbiotech Cat#19701 RNase Inhibitor

*  Other inhibitors: uridine 2¡¯, 3¡¯-cyclic vanadate, 5¡¯-diphosphoadeosine 3¡¯-phosphate and 5¡¯-diphosphoadeosine 2¡¯-phosphate (2), SDS, diethyl pyrocarbonate, 4M guanidinium thyocyanate plus 0.1M 2-Mercaptoethanol and heavy metal ions. Inactivated by phenol/chloroform extraction.

*  Inactivated by phenol/chloroform extraction.

*  Inactivated by heating at 95¡É for 10 minutes.

*  The working concentration for RNase A is 1-100 ug/ml depending on the application.

*  The enzyme is active under a wide range of reaction conditions. At low salt concentrations (0 to 100 mM NaCl), RNase A cleaves single-stranded and double-stranded RNA as well the RNA strand in RNA-DNA hybrids. However at NaCl concentrations of 0.3M or higher RNase A specifically cleaves single-stranded RNA.

 

If the troubleshooting guide does not solve the difficulty you are experiencing, please contact Technical Support with details of reaction setup, cycling conditions and relevant date.