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RNase A (DNase free)

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 ssRNA. RNase A is free of detectable DNase and protease activity, a heat treatment of the enzyme is not necessary before use.

Product Specifications

Product Name Alternative

Endoribonuclease, Bovine pancreas

Purity

≥ 90% (ion exchange chromatography), salt free, chromatographically homogeneous lyophilisate

Bioactivity

≥ 80 Kunitz units/mg

Form

Dry powder

Molecular Weight

Theoretical MW: 13.7 kDa (monomer)

Storage Conditions

Store at 2-8 °C. stock solutions in TE buffer should be aliquoted and stored at -20 °C

Notes

For research use only.

Applications Notes

Plasmid and genomic DNA preparation Removal of RNA from recombinant protein preparations Ribonuclease protection assays Mapping single-base mutations in DNA or RNA. Working concentration: 1-100 μg/ml (depending on application) . The enzyme is active under a wide range of reaction conditions. At low salt concentrations (0 to 100 mM NaCl), RNase cleaves ss and dsRNA as well as the RNA strand in RNA-DNA hybrids. At NaCl concentrations of 0.3 M or higher, RNase A specifically cleaves ssRNA. Stability: RNase A is an extremely stable enzyme, remarkable resistant to heating. It renatures easily after treatment with most denaturing agents. Isoelectric point (pI) : 9.6. Optimal pH: 7.0 (activity range 6-10) . Activity: ≥ 80 Kunitz units/mg. Unit definition: 1 Kunitz unit is that amount of activity which is capable of causing within 1 minute a decrease in absorbance at 300 nm equivalent to the maximum possible change in a 0.05% solution of yeast RNA at 25 °C, pH 5.0. Inactivation: Ribonuclease inhibitor, Vanadyl-ribonucleoside complexes, arabi-nonucleosides, Zn2+, Cu2+, penicillin, Vitamin B12, SDS, DEPC, 4 M guanidinium thiocyanate plus 0.1 M 2-mercaptoethanol. Most polyan-ions show some inhibitory effect. Inactivated by phenol/chloroform extraction.

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