Active Recombinant E. coli RNase H

• Cat.No.: rnh-63E
• Product Overview: A recombinant?E. coli?strain carrying the RNAse H (rnh) gene from?E.coli.

Specification

• Species: E.coli
• Source: E.coli
• Tag: Non
• Description: E.coli?RNase H (rnh) is an endoribonuclease which degrades the RNA strand of RNA/DNA hybrid molecules. RNase H digestion produces ribonucleotide molecules with 5-phosphate and 3-hydroxyl termini. RNAse H is nearly inactive against single or double-stranded RNA molecules.
• Bio-activity: 625,000 U/mg
• Purity: >99% by SDS-PAGE
• Unit Definition: 1 unit is defined as the amount of enzyme that will hydrolyze 1 nmol of RNA from an 3H-labeled DNA:RNA hybrid molecule into acid-soluble material in 20 minutes at 37°C.
• Storage: -25 to -15°C
• Concentration: 5,000 U/ml
• Storage Buffer: Supplied in: 20 mM Tris-HCl, 100 mM KCl, 10 mM MgCl2, 0.1 mM EDTA, 0.1 mM DTT, 50% glycerol, pH 7.9 @ 25°C Supplied with: 10X RNAse H Buffer: 500mM Tris-HCl, 750 mM KCl, 30 mM MgCl2, 100 mM DTT, pH 8.3 @ 25?C.

Citation

Characterization of Escherichia coli RNase H Discrimination of DNA Phosphorothioate Stereoisomers

Journal: Nucleic Acid Therapeutics    PubMed ID: 34619060    Data: 2021/12/1

Authors: ?ukasz J. Kie?piński, Erik Daa Funder, Peter H. Hagedorn

Article Snippet:Gel electrophoresis was performed in 1 × TBE buffer on 15% TBE-Urea gel (Novex ? ) with constant 180 V. Bands were visualized using ChemiDoc Touch Imaging System (Bio-Rad) on Blue Sample Tray.Gel electrophoresis was performed in 1 × TBE buffer on 15% TBE-Urea gel (Novex ? ) with constant 180 V. Bands were visualized using ChemiDoc Touch Imaging System (Bio-Rad) on Blue Sample Tray.. The recombinant E. coli RNase H enzyme used in the experiments was purchased from Creative BioMart (cat. RNASEH1-433H).. It was provided as a solution between 20 and 60 U/μL, but for the concentrations reported below, it was assumed to be 60 U/μL.It was provided as a solution between 20 and 60 U/μL, but for the concentrations reported below, it was assumed to be 60 U/μL.

Characterization of RNase H interaction with stereodefined PS backbone. (A) Structure of nucleic acid molecules used in the experiment. Base sequence of oligonucleotides in a box is a reverse complement of a sequence of RNA-containing molecule below the box. (B–D) Denaturing gel electrophoresis of RNA hydrolysis reactions.

Control of RNase H cleavage site with PS stereoconfiguration. (A) Structure of nucleic acid molecules used in the experiment. Arrows indicate major RNase H cleavage sites on complementary RNA molecule. (B) Denaturing gel electrophoresis of RNA hydrolysis reactions; “*”: full-length product, “#”: internal positive control cleavage site, “@”: cleavage induced by Rp-PS group within a stretch of 11 thymidines. Numbers above lanes indicate used oligonucleotide. Sample “5*” is a “no enzyme” control.

Massive parallel screen for preferred chiral motifs. (A) Structure of a circular substrate used in the experiment, with drawing of one of the possible RNase H interaction modes. (B) Denaturing gel electrophoresis showing hydrolysis of libraries upon treatment with RNase H. (C) Encoding of chiral status by nucleotide identity in different libraries. For the analysis, only the trinucleotides composed exclusively of A or T nucleotides in the randomized region were considered. (D) Changes in abundance of chiral motifs at different positions of the randomized gap of the circular substrate upon treatment with RNase H. Encoding 1: adenosine encodes Rp-PS, thymidine encodes Sp-PS; Encoding 2: opposite to encoding 1. Chiral motifs highlighted in red have the same effect direction and false discovery rate (FDR) below 0.01 in both of the compared libraries; highlighted in blue : as with red , but FDR <0.05. PS, phosphorothioate.