RNase A MBG

For removal of RNA during the isolation procedures of plasmid and genomic DNA

S_1358_6_LS_OEM_RNase_A_MBG_10_mg
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RNase A MBG (10 mg)

Cat. No. / ID:  RP14

Supplied in a salt-free, freeze-dried form Keep at -20°C (lyophilized or in a 50% glycerol solution) for long-term storage or at 4°C for up to several weeks.
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54,60 $
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Quantity
10 mg
50 mg
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The RNase A MBG is intended for molecular biology applications. This product is neither intended for the diagnosis, prevention or treatment of a disease, nor has it been validated for such use either alone or in combination with other products.
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Features

  • Possesses enzyme activity of >80 units/mg protein
  • Degrades RNA to cyclic nucleotide monophosphates to 5’-OH and 2’-, 3’-cyclic monophosphate
  • No endonuclease or exonuclease activity towards DNA substrates
  • Selectively cleaves single-stranded RNA 3’ next to pyrimidine residues

Product Details

The Ribonuclease A (RNase A) is a 13.7 kDa (monomer) endoribonuclease isolated from bovine pancreas, which selectively cleaves single-stranded RNA 3’ next to pyrimidine residues (cytosine, uracil). It degrades RNA to cyclic nucleotide monophosphates to 5’- OH and 2’-, 3’-cyclic monophosphate. The enzyme exhibits no endonuclease or exonuclease activity toward DNA substrates. RNase A removes RNA during the isolation procedures of plasmid and genomic DNA.

 

It is supplied with 1–10 mg/mL by resuspending in 10 mM Tris-HCl (pH 7.5), 15 mM NaCl, 50% (v/v) glycerol or in TE buffer.

 

One unit of activity is defined as the amount of enzyme which causes the hydrolysis of RNA to yield a velocity constant, k=1, at 25°C and pH 5.0.

Performance

Assay Specification
Activity 94.5 U/mg (Kunitz)
DNase contamination None detected
Protease contamination None detected

Principle

RNase A is very active under a wide range of reaction conditions and is difficult to inactivate. At low salt concentrations (up to 100 mM NaCl), the RNase A cleaves single- and double-stranded RNA as well as an RNA strand in RNA-DNA hybrids. However, under high salt concentrations (>300 mM NaCl), RNase A specifically cleaves single-stranded RNA.

 

RNase A cleaves specifically at the 3′-side of pyrimidine (uracil or cytosine) phosphate bonds. Ribonucleases do not hydrolyze DNA because the DNA lacks 2′-OH groups essential for forming cyclic intermediates. 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. RNase is supplied as a lyophilized powder.

Procedure

Usage

Stock solutions should be prepared to a final concentration of 1–10 mg/mL by resuspending in 10 mM Tris-HCl (pH 7.5), 15 mM NaCl, 50% (v/v) glycerol or in TE buffer. 

 

The recommended working solution concentration depends on the application.

 

  • For removal of RNA from plasmid preparations, use 10 μg/mL working solution and incubate the sample for 1 hour at room temperature.
  • For the preparation of „blunt ends” of double-stranded cDNA, use a 100 ng/mL working solution.

Applications

This is used for applications such as:

  • Purification of RNA-free DNA
  • Isolation of plasmid and genomic DNA
  • Removal of RNA during recombinant protein preparations
  • Protection of RNA during assays
  • Mapping of single-based mutations in DNA or RNA

 

Resources

Quick-Start Protocols (1)
Kit Handbooks (1)
RNase A MBG Manual
PDF (558KB)
Safety Data Sheets (1)
Certificates of Analysis (1)