RNase T1

- Not inactivated at 80°C in 20 min
- Recombinant enzyme
- LO certified

Catalog#	Size, concentration	Certificate of Analysis	MSDS

EN0541	100,000 u (1000 u/µl)	EN0541
EN0542	500,000 u (1000 u/µl)	EN0542

Product information

Applications

Removal of RNA from DNA preparations.
RNA sequencing (1).
Ribonuclease protection assays. Used in conjunction with RNase A (2).
Removal of RNA from recombinant protein preparations.
Determination of the level of RNA transcripts synthesized in vitro from DNA templates containing a "G-less cassette" (3).

Description

RNase T1 is an endoribonuclease that specifically degrades single-stranded RNA at G residues, see Fig.1. It cleaves the phosphodiester bond between 3'-guanylic residue and the 5'-OH residue of adjacent nucleotide with the formation of corresponding intermediate 2', 3'-cyclic phosphate (1). The reaction products are 3'-GMP and oligonucleotides with a terminal 3'-GMP.

RNase T1 does not require metal ions for activity.

Source

E.coli cells with a cloned rntA gene of Aspergillus oryzae.

Molecular Weight

11.2 kDa monomer.

Definition of Activity Unit

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.

Enzyme activity is assayed in the following mixture: 50 mM Tris-HCl (pH 7.5), 2 mM EDTA, 3 mg/ml yeast RNA.

Storage Buffer

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

Quality Control

The absence of endo-, exodeoxyribonucleases and proteases confirmed by appropriate quality tests. Functionally tested for RNA digestion in the plasmid DNA purification procedure (in conjunction with RNase A).

Inhibition and Inactivation

Inhibitors: 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 (4).
Inactivation by heating is reversible, reliably removed by spin column or phenol/chloroform extraction.

Figure 1. RNase T1 activity.

Patents, Licenses, Trademarks

Protocols & recommendations

RECOMMENDATIONS FOR USE

References

Takahashi, K., Moore, S., Ribonuclease T1, The Enzymes, V, (Boyer, P.D, Ed.), Academic Press, New York, the third edition, vol. 15, 435-468, 1982.
Sambrook, J., Russell, D.W., Molecular Cloning: A Laboratory Manual, the third edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 7.63-7.74, 2001.
Sawadogo, M., Roeder, R.G., Factors involved in specific transcription by human RNA polymerase II: Analysis by a rapid and quantitative in vitro assay, Proc. Natl. Acad. Sci. USA, 82, 4394-4398, 1985.
Eun, H-M., Enzymology Primer for Recombinant DNA Technology, Academic Press, Inc., 1996.

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