RNase T1 (C008B-269929)

RNase T1 is an endoribonuclease derived from Aspergillus oryzae. It specifically cleaves single-stranded RNA after guanosine residues (G), generating 3'-phosphate termini. RNase T1 forms nucleoside 2',3'-cyclic phosphate intermediates, cleaving the phosphodiester bond between the 3'-guanosine residue and the adjacent nucleoside 5'-OH group to produce oligonucleotides terminating in 3'-GMP and 3'-GMP. The activity of RNase T1 is independent of metal ions.Components ListR1522664ComponentsAppearance2KU5×2KUStorageR1522664ARNase T1 (500 U/μl)Liquid200ul5×200µl-20℃. Avoid freeze/thaw cycle.R1522664B10× RNase T1 BufferLiquid1ml5×1ml-20℃. Avoid freeze/thaw cycle.Unit DefinitionOne unit (U) is defined as the amount of enzyme required to produce an absorbance change of 1.0 at A260 using yeast RNA as the substrate at 37 °C and pH 7.5. ApplicationsRemoval of RNA contaminant from DNA preparations.Removal of RNA contaminant from recombinant protein preparations.RNA sequencing.Use in ribonuclease protection assays (in combination with RNase A).Quantitation of transcription levels from DNA templates with no or low guanine content.Quality ControlProtein PuritySDS-PAGE analysis confirms protein purity of no less than 95%.Non-specific Endonuclease Activity500 U of RNase T1 was incubated with 200 ng supercoiled plasmid DNA in a 20 μL reaction system at 37 °C for 4 hours. Agarose gel electrophoresis showed less than 20% of the plasmid DNA was converted to nicked or linear form.DNase Activity500 U of RNase T1 was incubated with 15 ng double-stranded DNA fragments in a 20 μL reaction system at 37 °C for 16 hours. Agarose gel electrophoresis showed no degradation of the dsDNA fragments.Usage Protocol1. Reaction systemComponentVolumeFinal ConcentrationssRNA1~10 μg50~500 ng/μL10× RNase T1 Buffer2 μL1×RNase Inhibitor (40 U/μL)*1 μL2 U/μLRNase T1 (500 U/μL)0.4 μL10 U/μLNuclease-Free Waterup to 20 μL-Note: To prevent degradation of the substrate RNA caused by contamination with environmental RNases such as RNase A, it is recommended to add RNase Inhibitor (Cat.No.: R350859) to the reaction system. The substrate RNA should be added only after the RNase Inhibitor has been added and thoroughly mixed.2. Recommended reaction conditions: Incubate at 37 °C for 15 minutes. The incubation time can be appropriately extended or shortened according to actual conditions to achieve optimal digestion results.NotesThe activity of RNase T1 can be inhibited by various metal ions, including Mg²⁺ (100 mM MgCl₂ inhibits approximately 40% of activity), Ca²⁺ (10 mM CaCl₂ inhibits approximately 30% of activity), Zn²⁺, Fe²⁺, Cu²⁺, etc. In addition, Guanylyl 2′-5′ guanosine is a specific inhibitor of RNase T1.Heat inactivation of RNase T1 is reversible. It is recommended to remove RNase T1 by column purification or phenol/chloroform extraction.RNase T1 is stable when heated at 100℃ for 10 min in solutions at pH 6.0, but unstable in alkaline solutions at pH > 9.0.For your safety and health, please wear a lab coat, disposable gloves, and a face mask during the experiment.

Specifications & Purity: RNase T1 is an endoribonuclease derived from Aspergillus oryzae. It specifically cleaves single-stranded RNA after guanosine residues (G), generating 3'-phosphate termini. RNase T1 forms nucleoside 2',3'-cyclic phosphate intermediates, cleaving the phosphodiester bond between the 3'-guanosine residue and the adjacent nucleoside 5'-OH group to produce oligonucleotides terminating in 3'-GMP and 3'-GMP. The activity of RNase T1 is independent of metal ions.Components ListR1522664ComponentsAppearance2KU5×2KUStorageR1522664ARNase T1 (500 U/μl)Liquid200ul5×200µl-20℃. Avoid freeze/thaw cycle.R1522664B10× RNase T1 BufferLiquid1ml5×1ml-20℃. Avoid freeze/thaw cycle.Unit DefinitionOne unit (U) is defined as the amount of enzyme required to produce an absorbance change of 1.0 at A260 using yeast RNA as the substrate at 37 °C and pH 7.5. ApplicationsRemoval of RNA contaminant from DNA preparations.Removal of RNA contaminant from recombinant protein preparations.RNA sequencing.Use in ribonuclease protection assays (in combination with RNase A).Quantitation of transcription levels from DNA templates with no or low guanine content.Quality ControlProtein PuritySDS-PAGE analysis confirms protein purity of no less than 95%.Non-specific Endonuclease Activity500 U of RNase T1 was incubated with 200 ng supercoiled plasmid DNA in a 20 μL reaction system at 37 °C for 4 hours. Agarose gel electrophoresis showed less than 20% of the plasmid DNA was converted to nicked or linear form.DNase Activity500 U of RNase T1 was incubated with 15 ng double-stranded DNA fragments in a 20 μL reaction system at 37 °C for 16 hours. Agarose gel electrophoresis showed no degradation of the dsDNA fragments.Usage Protocol1. Reaction systemComponentVolumeFinal ConcentrationssRNA1~10 μg50~500 ng/μL10× RNase T1 Buffer2 μL1×RNase Inhibitor (40 U/μL)*1 μL2 U/μLRNase T1 (500 U/μL)0.4 μL10 U/μLNuclease-Free Waterup to 20 μL-Note: To prevent degradation of the substrate RNA caused by contamination with environmental RNases such as RNase A, it is recommended to add RNase Inhibitor (Cat.No.: R350859) to the reaction system. The substrate RNA should be added only after the RNase Inhibitor has been added and thoroughly mixed.2. Recommended reaction conditions: Incubate at 37 °C for 15 minutes. The incubation time can be appropriately extended or shortened according to actual conditions to achieve optimal digestion results.NotesThe activity of RNase T1 can be inhibited by various metal ions, including Mg²⁺ (100 mM MgCl₂ inhibits approximately 40% of activity), Ca²⁺ (10 mM CaCl₂ inhibits approximately 30% of activity), Zn²⁺, Fe²⁺, Cu²⁺, etc. In addition, Guanylyl 2′-5′ guanosine is a specific inhibitor of RNase T1.Heat inactivation of RNase T1 is reversible. It is recommended to remove RNase T1 by column purification or phenol/chloroform extraction.RNase T1 is stable when heated at 100℃ for 10 min in solutions at pH 6.0, but unstable in alkaline solutions at pH > 9.0.For your safety and health, please wear a lab coat, disposable gloves, and a face mask during the experiment.

Grade: Recombinant;DNase free;Suitable for molecular biology;EnzymoPure™

Molecular Weight: 11 kDa

EC Number: 232-809-1

Storage Temperature: Store at -20°C;Avoid repeated freezing and thawing

Shipped in: Ice chest + Ice pads

UPC:

12352204

Weight:

7.06 Ounces

HazmatClass:

Yes

Quantity:

1

Unit of Measurement:

EA

WeightUOM:

LB

MPN:

R1522664-100KU

Country of Origin:

China

CAS:

9026-12-4

Product Size:

100KU

Harmonized Tariff Code:

2931.90.90