RNase H (E.coli)

RNase H (E.coli) 

Cat. No.: SB-G3417-100U
Size: 100U


Description

This product RNase H, namely ribonuclease H, is derived from Escherichia coli and is an endoribonuclease. RNase H specifically hydrolyzes the RNA in the DNA-RNA hybrid strand, and the hydrolysis products are oligoribonucleotides and single-stranded DNA at the 5' phosphate end, and cannot hydrolyze single-stranded or double-stranded DNA or RNA. This product is mainly used to remove mRNA before the second strand of cDNA is synthesized, remove poly(A) in mRNA hybridized with poly(dT), and achieve fixed-point shearing of RNA through DNA sequence, etc.

• Source : derived from Escherichia coli, recombinantly expressed by Escherichia coli.
• Definition of enzyme activity : at 37°C, the amount of enzyme required to catalyze 1 nmol of RNA in the hybrid duplex to form acid-soluble ribonucleotides within 20 minutes is defined as one enzyme activity unit.
• Purity and concentration : SDS-PAGE detection purity ≥ 95%; 2 U/μL.
• Inactivation or inhibition : Incubation at 65°C for 20 minutes can inactivate; both metal ion chelating agents and sulfhydryl blocking agents can inhibit RNase H activity.
• Enzyme storage buffer : 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1 mM DTT, 200μg/mL BSA, 50% Glycerol, pH 7.4.
• 10 × Reaction Buffer : 500 mM Tris-HCl, 30 mM MgCl₂ , 10 mM DTT, pH 8.3.

Storage and Handling Conditions

1. After using reverse transcriptase to synthesize the first strand of cDNA, incubate at 70°C for 10 minutes to terminate the reaction;
2. Add 2 μL 10х Reaction buffer, appropriate amount of Nuclease-Free water and 0.5-1 μL RNase H to the 20 μL reaction system on an ice bath, and mix well;
3. Incubate the reaction system at 37°C for 1 h;
4. After the reaction is completed, incubate at 65°C for 20 min to terminate the reaction.

1. All enzymes should be stored in an ice box and stored at -20°C immediately after use.
2. For your safety and health, please wear a lab coat and disposable gloves for operation.