Exonuclease III (E.coli), 5000 U (for Nuclease/Topoisomerase)

Product Information

Product Introduction

Exonuclease III ( E. coli ) is an exonuclease III derived from Escherichia coli, expressed recombinantly in E. coli, and exhibits four catalytic activities. The 3'-5' deoxyribonuclease activity is specific for double-stranded DNA, and gradually cuts off the 5'-single nucleotide from the 3'OH terminal. Its best substrate is blunt-end, 5' overhang or nicked double-stranded DNA. It has no activity on DNA 3'-overhang (at least 4 bases long and without a 3'-terminal C-residue), single-stranded DNA or thiophosphate-linked nucleotides. Exonuclease III ( E. coli ) has 3' phosphatase activity that can remove the 3'-terminal phosphate group and generate a 3'-OH group. Its RNase H activity can degrade the RNA chain in RNA-DNA hybrids in an exonuclease manner. The apurinic/apyrimidinic-endonuclease activity cleaves the phosphodiester bond at the apurinic or apyrimidinic site to produce a 5'-end (abasic deoxyribose 5'-phosphate residue). It is mainly used for single-stranded template preparation, site-directed mutagenesis, and strand-specific probe preparation for DNA dideoxy sequencing.

Source: Derived from Escherichia coli and expressed by recombinant Escherichia coli.

Definition of enzyme activity: The amount of enzyme required to produce 1 nmol of acid-soluble substance within 30 min at 37°C and pH 8.0 is defined as one unit of enzyme activity.

Purity and concentration: Purity detected by SDS-PAGE 95%; 100 U/μL.

Inactivation: Incubate at 70 for 20 min to inactivate

Enzyme storage buffer: 25 mM Tris-HCl, 50 mM KCl, 0.5 mM DTT, 50% Glycerol, pH 8.0.

10х Reaction buffer：500 mM Tri-HCl，100 mM MgCl₂，10 mM DTT，pH 8.0。

Storage and transportation

Transported with wet ice; stored at -20, valid for 12 months.

composition

Procedure

1. Configure the reaction system according to the following table:

2. Mix the reaction system thoroughly and incubate at 37°C for 30 min.

3. After the reaction is completed, incubate at 70 for 20 min to terminate the reaction. Then proceed with subsequent experimental operations.

Precautions

1. The enzyme should be placed in an ice box when used and stored at -20 immediately after use.

2. For your safety and health, please wear a lab coat and disposable gloves when operating.

The product is for scientific research purposes only and not for clinical diagnosis!

https://www.servicebio.com/goodsdetail?id=12056