Exonuclease III (E. coli)
Cat. No.: SB-G3465-5000U
Size: 5000U
Description
Exonuclease III ( E.coli ) is an exonuclease III derived from Escherichia coli, which is recombinantly expressed in Escherichia coli and exhibits four catalytic activities. The 3'-5' exodeoxyribonuclease activity is specific to double-stranded DNA, and the 5'-mononucleotide is gradually cut from the direction of the 3'OH end, and the best substrate is a blunt end and a 5' overhang or nicked double-stranded DNA. It is not active on DNA 3'-overhangs (at least 4 bases long and without a 3'-terminal C-residue), single-stranded DNA, or phosphorothioate-linked nucleotides. Exonuclease III ( E.coli ) has a 3' phosphatase activity that removes the 3'-terminal phosphate group to generate a 3'-OH group. Its RNase H activity can degrade RNA strands in RNA-DNA hybrids in an exonuclease manner. Apurinic/apyrimidinic-endonuclease activity cleaves phosphodiester bonds at apurinic or apyrimidinic sites, generating 5'-terminals (abasic deoxyribose 5'-phosphate residues). It is mainly used for single-strand template preparation, site-directed mutagenesis, and strand-specific probe preparation for DNA dideoxy sequencing.
- Source : derived from Escherichia coli, recombinantly expressed by Escherichia coli.
- Definition of enzyme activity : at 37°C and pH 8.0, the amount of enzyme required to produce 10 nmol of acid-soluble substances within 30 minutes is defined as one enzyme activity unit.
- Purity and concentration : SDS-PAGE detection purity ≥ 95%; 100 U/μL.
- Inactivation or inhibition : Incubation at 70°C for 20 min can inactivate.
- Enzyme storage buffer : 25 mM Tris-HCl, 50 mM KCl, 0.5 mM DTT, 50% Glycerol, pH 8.0.
- 10 x Reaction Buffer : 500 mM Tris-HCl, 10 mM DTT, 100 mM MgCl2 , pH 8.0.
Storage and Handling Conditions
Transport in wet ice packs; store at -20°C, valid for 12 months.
Composition
| Component Number |
Component |
G3465 |
| G3465-1 |
Exonuclease III ( E. coli ) |
50 μL |
| G3465-2 |
10 × Reaction Buffers |
500 μL |
| Product Manual |
1 copy |
Steps
1. Configure the reaction system according to the following table:
| Component |
Volume |
| Exonuclease III ( E. coli ) |
0.5 μL |
| 10 × Reaction Buffers |
5 μL |
| DNA sample |
≤5μg |
| Nuclease Free Water |
To 50 μL |
2. Mix the reaction system well and incubate at 37°C for 30 min.
3. After the reaction is complete, incubate at 70°C for 20 min to terminate the reaction. Subsequent experimental operations can be carried out.
Precautions
- All enzymes should be stored in an ice box and stored at -20°C immediately after use.
- For your safety and health, please wear a lab coat and disposable gloves for operation.
