Medical Laboratory Reagents
|CAS No. 9001-99-4 EC No. 22.214.171.124
|From Eukaryotic cells(Yeast) with cloned gene encoding genetically engineered Bovine pancreatic ribonuclease
||Ribonuclease A, Pancreatic Ribonuclease, Ribonuclease I, Ribonuclease 3’-pyrimidinooligonucleotidohydrolase
optimal pH 7.6
||4 °C for lyophilized powder
-20 °C for solution
||maximum activity at 60 °C
|Temperature range of enzyme activity:
|| Maximum activity observed at 60 °C, although the enzyme exhibit activity in the temperature range of 15-70 °C.
||= 60 Kunitz units/mg of protein
| Unit definition:
||One unit of the enzyme causes an increase of absorbance by 1.0 unit at 260 nm when yeast RNA is hydrolyzed at 37°C and pH 5.0.
Fifty units are approximately equivalent to 1 Kunitz unit.
||E1% = 7.1 (280 nm)
|Purity and Quality
||Molecular biology grade
Endonuclease and exonuclease none detected
Free of DNase activity. No need to heat it before use.
| General Description
|| RNase A is an endoribonuclease that specifically cleaves the single strand RNA at the phosphodiester bond between the 5’-ribose of a nucleotide and the 3’ OH phosphate group of an adjacent pyrimidine nucleotide.
The highest activity is demonstrated with single stranded RNA.
||The recombinant enzyme is identical to the native RNase A in amino acid sequence, molecule structure and specifications. However, the recombinant preparation has advantage in lot-to-lot consistency, superior purity and cost-efficiency.
|| RNase A is used to remove RNA from plasmid and genomic DNA preparation. It also used to remove RNA from protein samples.
RNase protection assays
Analysis of RNA sequences
Mapping single-base mutations in DNA or RNA
| Preparation Notes
|| Potassium and sodium salts
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 0.3 M or higher, RNase A specifically cleaves single-stranded RNA
|| By alkylation of His 12 and His 119
The RNase A has a high affinity to glass surfaces.
At neutral pH and high concentrations (>10 mg/ml) the enzyme will precipitate.
The enzyme is inhibited by diethyl pyrocarbonate (DEPC), guanidinium salts (4 M GuSCN), ß-mercaptoethanol, heavy metals and RNase-inhibitors like RIBOPROTECT (RT33).
In order to remove the enzyme from a sample, perform a separation with spin columns or several phenol/chloroform extractions.
| Preparation Instructions
|| Recommended concentration of RNase A is 1 to 100 µg/mL depending on the application.
For removal of RNA from plasmid preparations use 10 µg/ml working solution and incubate sample for 1 hour at room temperature.
For preparation of „blunt ends” of double-stranded cDNA use 100 ng/ml working solution.
||storage at -20 °C
recommended storage at -20 °C for solution and lyophilized powder to maintain stable for at least 2 years.
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
|Precautions and Disclaimer
|| This product is for R&D use only, not for drug, household, or other uses.
RNase A is stable to both heat and detergents. In addition, it adsorbs strongly to glass. Scrupulous precautions are necessary to ensure RNase A residue does not cause artifacts in processes requiring intact RNA.