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RNase T1

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

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

RNase T1 does not require metal ions for activity.

Applications

  • Removal of RNA from DNA preparations
  • RNA sequencing (see​ Reference 1)
  • Ribonuclease protection assays. Used in conjunction with RNase A (see​ Reference 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" (see​ Reference 3)
  
Storage Condition-20 C
HazardousNo
Concentration1000 U/µL
Definition of Activity UnitOne 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.
InactivationInactivation by heating is reversible, reliably removed by spin column or phenol/chloroform extraction.
InhibitionInhibitors: 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).
Molecular Weight11.2 kDa monomer.
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).
SourceE. 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.
RNase T1 activity

RNase T1 activity

RNase T1 activity

RNase T1 activity.


References

  1. K. Takahashi, S. Moore, Ribonuclease T1, The Enzymes, V (Boyer, P.D, Ed. 3, Academic Press, New York, the third edition, vol. 15, 1982), pp. 435-468.
  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), pp. 7.63-7.74.
  3. M. Sawadogo, R. G. Roeder, 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).
  4. H-M. Eun, Enzymology Primer for Recombinant DNA Technology (Academic Press, Inc., 1996).

Protocols and Product Manuals

pdf   RNase T1 - Product Information