Exonuclease I (E. coli)
Cat. No.: SB-G3464-3000U
Size: 3000U
Description
Exonuclease I ( E.coli ) is an exonuclease I derived from Escherichia coli, which is recombinantly expressed in Escherichia coli, degrades single-stranded DNA in the 3'-5' direction, and decomposes to generate 5'-mononucleotides . It has very high specificity for single-stranded DNA and cannot decompose double-stranded DNA and RNA. It is mainly used to remove residual primers after PCR reaction or remove ssDNA fragments in the reaction system.
- Source : derived from Escherichia coli, recombinantly expressed by Escherichia coli.
- Definition of enzyme activity : at 37°C and pH 9.5, 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%; 20 U/μL.
- Inactivation or inhibition : Incubation at 80°C for 20 min can inactivate.
- Enzyme storage buffer : 20 mM Tris-HCl, 0.5 mM EDTA, 1 mM DTT, 50% glycerol, pH 7.5.
- 10 x Reaction Buffer : 670 mM Glycine-KOH, 10 mM DTT, 67 mM MgCl2 , pH 9.5.
Storage and Handling Conditions
Transport in wet ice packs; store at -20°C, valid for 12 months.
Composition
| Component Number |
Component |
G3464 |
| G3464-1 |
Exonuclease I ( E. coli ) |
150 μL |
| G3464-2 |
10 × Reaction Buffers |
500 μL |
| Product Manual |
1 copy |
Steps
1. Configure the reaction system according to the following table:
| Component |
Volume |
| Exonuclease I ( E. coli ) |
1 μL |
| 10 × Reaction Buffers |
2 μL |
| DNA sample |
x μL |
| Nuclease Free Water |
To 20 μL |
2. Mix the system thoroughly and incubate at 37°C for 30 minutes to remove single-stranded DNA in the system. (Exonuclease I can also be added directly to the PCR product)
3. After the reaction is complete, incubate at 80°C for 20 min to terminate the reaction.
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.
