RNase T1

Item Number

R749974

• Specifications & Purity: ≥95%, free of DNA endonuclease and exonuclease, and other ribonucleases.
• Stability And Storage: Store at -20 ℃ for up to 2 years.
• Storage Temp: Store at -20°C
• Shipped In: Ice chest + Ice pads

Product Description

One unit of the enzyme causes an increase in absorbance of 1.0 at 260nm in 15min when yeast RNA is hydrolyzed at 37℃ and pH7.5.

Application：

RNA removal from DNA; RNA removal from recombinant proteins; RNA sequencing; RNA protection assay; detecting the transcription level of G-free or low-G DNA templates.

Source：

Aspergillus oryzae RNase T1 expressed in E, coli.

Enzyme storage buffer：

50mM Tris-HCl (pH7.4), 50% (v/v) glycerol.

Inactivation or inhibition：

Metal ions Mg2+ (100mM MgCl2 inhibits about 40% activity), Ca2+ (10mM CaCl2 inhibits about 30% activity), Zn2+, Fe2+, Cu2+ and single nucleotides (2'- GMP, 3' -GMP, etc.) can inhibit the activity of RNase T1. Guanylyl 2'-5' guanosine is a specific inhibitor of RNase T1 [2]. RNase T1 is highly tolerant to heat. Under pH 6.0, it can tolerate 100℃ for 10 minutes, but it is unstable in alkaline solutions with a pH ＞ 9.0. RNase T1 after heat-inactivation is reversible. The reaction can not be terminated by incubation at 100℃ [3]. RNase T1 can be removed by column purification or phenol/chloroform extraction.

Precautions：

Keep RNase T1 on ice when handing, and store at -20 ℃ immediately after use.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation.

Instructions for Use：

1. Digestion of single-stranded RNA containing Ga. Set up the reaction on ice as follows:ReagentVolumeFinal ConcentrationDEPC Water (DNase, RNase free)(16.4-x)μl-10X Reaction Buffer2μl1XRNase Inhibitor (40U/μl)1μl2U/μlDTT (100mM)0.4μl2mMSingle-stranded RNA Containing G NucleotidexμlRNA≤50ng/μlRNase T1 (1000U/μl)0.2μl10U/μlTotal volume20μl-Note 1: When multiple reactions are required, prepare a master mix including all reagents except the substrate RNA and then dispense to different nuclease-free tubes. Finally, add the substrate RNA into each tube.Note 2: To prevent RNA degradation, it is recommended to add RNase Inhibitor into the reaction mixture (in order to maintain the activity of RNase Inhibitor, the solution needs to contain at least 1mM DTT). Substrate RNA should be added after the addition of RNase Inhibitor.a. Mix well by pipetting gently. Centrifuge briefly to collect liquid at the bottom of the centrifuge tube. b. Incubate at 37℃ for 15 minutes. The incubation time can be appropriately extended to ensure a sufficient digestion. Usually, for 1μg of RNA in a 20μl reaction volume with 0.2μl of RNase T1 (1000U/μl), it is recommended to incubate for 30 minutes. If the amount of substrate RNA is relatively small, the incubation time can be appropriately reduced.2. For other applications, please refer to the protocol above or other relevant literature.References:1. Takahashi K, Moore S. The Enzymes, V. Academic Press. 1982. 435-468.2. Eun H-M. Enzymology Primer for Recombinant DNA Technology. Academic Press. 1996.3. Uchida T, Egami F. The Enzymes, IV. Academic Press. 1971. 205-250.