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Cat. No.: RNT1-100k (for 100KU)

Cat. No.: RNT1-500k (for 500KU)

Description

RNase T1 is derived from Aspergillus oryzae and is a ribonuclease, specifically cleaving after guanosine (G) residues in single-stranded RNA. During the reaction, RNase T1 cleaves the phosphodiester bond between the guanosine nucleotide's 3'-phosphate group and the adjacent nucleotide's 5'-hydroxyl group, resulting in a 2',3'-cyclic phosphoadenosine intermediate. Even if a G-cP (guanosine-3'-cyclic phosphate) is formed at the 3' end, it is subsequently hydrolyzed to produce an oligonucleotide with GMP (G-P) at the 3' end. The activity of RNase T1 is independent of metal ions.

Applications

• Removal of RNA from DNA preparations.
• Removal of RNA from recombinant protein preparations.
• RNA sequencing.
• Used in combination with RNase A for RNA protection analysis (RNA protection assay).
• Determination of transcription levels in DNA templates lacking G or with low G content.

Source

The enzyme is expressed in Escherichia coli, and the expressed gene is rntA from Aspergillus oryzae.

Molecular Weight

12.4 kDa.

Activity Definition

One unit of the enzyme causes an increase in absorbance of 1.0 at 260nm in 15 minutes when yeast RNA is hydrolyzed at 37°C and pH 7.5.

Purity

Purity is ≥95%; it does not contain endonucleases or exonucleases, and there are no other ribonucleases present.

Enzyme Storage Solution

50mM Tris-HCl (pH 7.4), 50% (v/v) glycerol.

Reaction Buffer (10X)

500mM Tris-HCl (pH 7.5), 20mM EDTA.

Inactivation or Inhibition

Metal ions Mg2+ (approximately 40% inhibition at 100mM MgCl2), Ca2+ (approximately 30% inhibition at 10mM CaCl2), Zn2+, Fe2+, Cu2+, and nucleotides (such as 2'-GMP, 3'-GMP) can inhibit RNase T1 activity. Guanylyl 2'-5' guanosine is a specific inhibitor of RNase T1. RNase T1 exhibits high thermal tolerance and can withstand heating at 100°C for 10 minutes in a pH 6.0 solution, but it is unstable in alkaline solutions with pH > 9.0. The thermal inactivation of RNase T1 is reversible. Heating the reaction system to 100°C does not terminate the reaction. RNase T1 can be removed by column purification or phenol/chloroform extraction methods.

Storage

Store at -20°C, valid for at least two years.

Precautions

• The enzyme should be stored in an icebox or on ice during use, and after use, it should be immediately placed at -20°C for storage.
• This product is intended for scientific research by professionals only and is not to be used for clinical diagnosis or treatment, food, or drugs. It must not be stored in residential homes.
• For your safety and health, please wear a lab coat and disposable gloves while handling.

Only for research and not intended for treatment of humans or animals

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