Uracil-DNA Glycosylase (UDG)
Cat. No.: SB-G3419-1000U
Size: 1000U
Description
Uracil-DNA Glycosylase, referred to as UDG, is derived from Escherichia coli and can catalyze the hydrolysis of the N-glycosidic bond between the uracil (dU) base and deoxyribose in the uracil-containing DNA chain, thereby releasing free uracil. Uracil-DNA Glycosylase (UDG) can hydrolyze single-stranded or double-stranded DNA containing dU, but cannot hydrolyze RNA or DNA oligomers containing dU with a length of no more than 6 bases. UDG is mainly used to eliminate the product contamination problem during PCR amplification.
- Source : derived from Escherichia coli, recombinantly expressed by Escherichia coli.
- Definition of enzyme activity : at 37°C, the amount of enzyme required to catalyze the release of 60 pmol of uracil from uracil-containing double-stranded DNA within 1 min is defined as one enzyme activity unit. Enzyme activity was determined by the amount of [ 3 H]-uracil released from a 50 μL reaction system containing 0.2 μg DNA (104 -105 cpm/μg) within 30 minutes .
- Purity and concentration : SDS-PAGE detection purity ≥ 95%; 5 U/μL.
- Inactivation: Incubating at 95°C for 10 minutes can inactivate 95% of UDG.
- Enzyme storage buffer : 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1 mM DTT, 0.1 mg/mL BSA, 50% glycerol, pH 7.4.
- 10 × Reaction Buffer : 200 mM Tris-HCl, 10 mM EDTA, 10 mM DTT, pH 8.0.
Storage and Handling Conditions
Transport in wet ice packs; store at -20°C, valid for 12 months.
Composition
| Component Number |
Component |
G3419-1000U |
| G3419-1 |
Uracil-DNA Glycosylase (UDG) |
200 μL |
| G3419-2 |
10 × UDG Reaction Buffers |
1 mL |
| Product Manual |
1 copy |
Steps
1. Configure the reaction system according to the following table:
| Component |
Volume |
| Uracil-DNA Glycosylase (UDG) |
0.2 μL |
| Taq DNA polymerase (5 U/μL) |
0.25 μL |
| template |
x μL |
| Primer Mix (10 μM each) |
4 μL |
| dNTP/dUTP (2.5 mM each/5 mM) |
4 μL |
| 10 × PCR Reaction Buffers |
5 μL |
| Nuclease Free Water |
To 50 μL |
2. After the reaction system is prepared, mix well and incubate at 37°C for 10 minutes (this step can effectively remove possible contamination of PCR products containing dUTP before).
3. Follow-up can immediately enter the PCR amplification program.
Precautions
- UDG enzyme is active in most PCR reaction buffer systems, but for self-used PCR or RT-PCR systems, it is recommended to test whether it is compatible with the system used for the first time.
- 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.
