Uracil-DNA Glycosylase (UDG)
Product Introduction
Uracil-DNA Glycosylase (UDG) is the first key enzyme in the DNA base excision repair (BER) pathway, playing an important role in maintaining genomic integrity and in molecular biology experiments.
UDG possesses highly specific substrate recognition capabilities, accurately identifying uracil bases in DNA strands resulting from dUTP incorporation or spontaneous deamination of cytosine. Its mechanism of action involves hydrolyzing the N-glycosidic bond between uracil and deoxyribose, releasing free uracil while simultaneously creating an abasic site (AP site) in the DNA backbone. This process requires no cofactors and exhibits extremely high catalytic efficiency. UDG shows no activity against uracil in RNA strands or other mismatched bases in double-stranded DNA, ensuring it acts only on the correct repair targets.
Product Specifications
Parameter | Specification |
Source | Recombinant expression in E. coli |
Molecular Weight | ~29 kDa |
Concentration | 5,000 units/ml |
Activity | 3'→5' single-stranded DNA exonuclease activity |
Purity | ≥95% (SDS-PAGE) |
Endotoxin | <1 EU/μg |
Storage Buffer | 10 mM Tris-HCl, 50 mM KCl, 0.5 mM EDTA, 1 mM DTT, 100 μg/ml BSA, 50% Glycerol, pH 7.5 |
10× Reaction Buffer | 200 mM Tris-HCl, 10 mM DTT, 10 mM EDTA, pH 8.0 |
Storage Conditions | Long-term storage at -80°C; short-term storage at -20°C |
Product Specifications
Specifications | Catalog Number | Concentration | Volume |
250 U | GR300201 | 5,000 U/mL | 50 μL |
1,250 U | GR300202 | 5,000 U/mL | 250 μL |
2,500 U | GR300203 | 5,000 U/mL | 500 μL |
Application Scenarios
PCR Contamination Control: The primary application of UDG in PCR experiments is to prevent aerosol contamination. By incorporating dUTP instead of dTTP into amplification products, UDG can selectively degrade dU-containing contaminants, while natural DNA templates (containing dT) remain unaffected. This technology is widely used in detection systems such as qPCR and RT-qPCR, significantly reducing the risk of false positives, particularly in clinical nucleic acid testing (e.g., for SARS-CoV-2).
USER Cloning (Uracil-Specific Excision Reagent): UDG, used in conjunction with Endonuclease VIII, can generate sticky ends at dU sites, simplifying the directional ligation of vectors and insert fragments. This is used for plasmid construction or TALE protein assembly.
Strand-Specific RNA Library Construction: By labeling the second strand of cDNA with dUTP, UDG treatment can selectively remove the non-target strand, preserving strand-specific information and improving the accuracy of transcriptome analysis.
References
Jacob LP, et al. (2021). The N-terminal domain of uracil-DNA glycosylase: Roles for disordered regions. DNA Repair (Amst). 101:103077.
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