DNA-Free Thermostable UDG (Ura

Thermostable UDG (Uracil-DNA Glycosylase) hydrolyzes the N-glycosidic bond between uracil bases and deoxyribose in DNA chains, releasing free uracil. This enzyme acts on single-stranded or double-stranded DNA containing dU and is not active against RNA. It is sourced from thermophilic microorganisms and exhibits higher enzymatic activity at elevated temperatures.

Explore Versatile Options with Our Glycerol-Free and Lyophilized Variants

Delight in the flexibility of our product offerings, now available in a glycerol-free version, meticulously crafted to seamlessly integrate with your freeze-drying systems. 

Additionally, we present a lyophilized version, a practical solution ensuring stable transportation at room temperature, thereby notably mitigating shipping costs, especially during long-distance transits.

Cat. No.: DFTSUNG-200 (for 200U)

Cat. No.: DFTSUNG-1k (for 1000U)

For the heat-labile version, please visit HL-UDG.

The lyophilized Thermostable UDG (Uracil-DNA Glycosylase) is also available by inquiry, which can be transported at room temperature.

Description

Thermostable UDG (Uracil-DNA Glycosylase) hydrolyzes the N-glycosidic bond between uracil bases and deoxyribose in DNA chains, releasing free uracil. This enzyme acts on single-stranded or double-stranded DNA containing dU and is not active against RNA. It is sourced from thermophilic microorganisms and exhibits higher enzymatic activity at elevated temperatures.

In high-temperature PCR reactions, especially during denaturation and extension phases, dCTP undergoes deamination to form dUTP. This dUTP can be incorporated into DNA by polymerases, resulting in the presence of dU in the DNA. Additionally, high temperatures can cause deamination of dC bases in DNA to form dU bases. These dU bases in DNA can inhibit the activity of high-fidelity DNA polymerases, such as Pfu DNA polymerase. Therefore, removing dU bases from DNA can enhance the amplification yield of high-fidelity DNA polymerases. Our high-temperature UDG enzyme can withstand temperatures up to 95°C, with a half-life exceeding 1 hour at 95°C. Thus, this high-temperature UDG serves as an excellent enhancer for increasing the amplification yield of high-fidelity DNA polymerases like Pfu DNA polymerase.

Our DNA-Free Thermostable UDG, on the other hand, is devoid of genomic and plasmid DNA from the production strain. This makes DNA-Free Thermostable UDG) particularly suitable for amplifying, sequencing, and cloning 16S and 23S rDNA genes.

Activity Definition

One unit of activity is defined as the amount of enzyme required to catalyze the release of 60 pmol of uracil from uracil-containing double-stranded DNA (dU/dA) per minute at 65°C.

Activity Assay Conditions

Pfu PCR Buffer: 20 mM Tris-HCl (pH 8.8 at 25°C), 10 mM (NH4)2SO4, 10 mM KCl, 0.1 mg/mL BSA, 0.1% (v/v) Triton X-100, 2 mM MgSO4, incubated at 65°C.

Features

• Heat-stable, retains enzymatic activity at temperatures ranging from 50-90°C.
• Active in a variety of PCR enzyme buffers.

Applications

• Elimination of dU generated at high temperatures in DNA, improving the performance of high-fidelity polymerases in amplification.
• Other experimental procedures requiring the removal of dU at high temperatures.

Storage

Store at -20°C.