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RNase A/T1 Mix

LO certified Not inactivated at 80°C in 20 min

RNase A/T1 Mix combines the RNA degradation activity of both RNase A and RNase T1.
  
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Thermo Scientific RNase A/T1 Mix combines the RNA degradation activity of both RNase A and RNase T1. The RNase A specifically hydrolyzes RNA at C and U residues; RNase T1 specifically hydrolyzes RNA at G residues (see Figure 1 in Supporting Data [see Reference 1]).

Highlights

  • Higher level of RNA degradation than with RNase A and RNase T1 separately
  • Free of DNase activity - It is not necessary to heat the mix before use

Applications

  • Removal of RNA from DNA preparations (see Reference 2)
  • Removal of RNA from recombinant protein preparations
  • Ribonuclease protection assays (see​ Reference 2)
  
Storage Condition-20 C
HazardousNo
Concentration2 mg/mL (approx. 10,000 U/mL (200 Kunitz U/mL)) of RNase A; 5000
InactivationNot inactivated by heating, reliably removed by spin column or phenol/chloroform extraction.
InhibitionInhibitors for RNase A: the most potent inhibitor is a mamalian ribonuclease inhibitor, e.g., RiboLock RNase Inhibitor. Other inhibitors: uridine 2', 3'-cyclic vanadate, 5'-diphosphoadenosine 3'-phosphate, and 5'-diphosphoadenosine 2-phosphate (see Reference 2), SDS, diethyl pyrocarbonate, 4 M guanidinium thiocyanate plus 0.1 M 2-mercaptoethanol and heavy metal ions.

Inhibitors for RNase T1: metal ions Mg2+, Ca2+, Zn2+, Fe2+, Cu2+ (MgCl2 at 100 mM concentration is approx. 40% inhibitory, CaCl2 at 10 mM is approx. 30% inhibitory); mononucleotides (2'-GMP, 3'-GMP, etc.); guanilyl-2',5'-guanosine is a specific inhibitor (see Reference 4).
Quality Control
  • The absence of endo-, exodeoxyribonucleases, and proteases confirmed by appropriate quality tests.
  • Functionally tested for degradation of RNA in the plasmid DNA isolation procedure.
SourceRNase A: Bovine pancreas.
RNase T1: E.coli cells with a cloned rntA gene of Aspergillus oryzae.
Storage BufferThe enzyme is supplied in 50 mM Tris-HCl (pH 7.4) and 50% (v/v) glycerol.
Unit DefinitionRNase A: One unit of the enzyme causes an increase in absorbance of 1.0 at 260 nm when yeast RNA is hydrolyzed at 37°C and pH 5.0.
Fifty units are approximately equivalent to 1 Kunitz unit (see Reference 3).

RNase T1: One unit of the enzyme causes an increase in absorbance of 1.0 at 260 nm in 15 min when yeast RNA is hydrolyzed at 37°C and pH 7.5.
RNase A/T1 Mix activity

RNase A/T1 Mix activity

RNase A/T1 Mix activity

RNase A/T1 Mix activity.


References

  1. F. M. Ausubel et al., Current Protocols in Molecular Biology, vol.1, (John Wiley).
  2. J. Sambrook, D. W. Russell, Molecular Cloning: A Laboratory Manual, the third edition (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 2001).
  3. M. A. Kunitz, A spectrophotometric method for the measurement of ribonuclease activity. J. Biol. Chem. 164, 563-568, (1946).

Protocols and Product Manuals

pdf   RNase A/T1 Mix - Product Information