Thermolabile Uracil-DNA Glycosylase (UNG) Enzyme - Highly Specific for dU Removal in PCR and NGS Sample Preparation, Enhancing DNA Quality Control

Description

Uracil-DNA Glycosylase (UNG), Thermolabile is a recombinant protein in E. coli, from marine bacterium. The enzyme hydrolyzes uracil-glycosidic bonds at U-DNA in single- and double-stranded DNA excising uracil and creating alkali sensitive abasic sites in the DNA. The enzyme is inactive on RNA and native, uracil-free DNA. Since Uracil-DNA Glycosylase (UNG), Thermolabile has no metal ion requirements, it is fully active in the presence of EDTA.

Uracil-DNA Glycosylase (UNG), Thermolabile can be used with dUTP to eliminate PCR “carry over” contaminations from previous DNA synthesis reactions. To make PCR products suspectible to degradation, dTTP has to be substituted by dUTP in the PCR reaction mix. Subsequent PCR reaction mixes must be pretreated with Uracil-DNA Glycosylase (UNG), Thermolabile prior to PCR to degrade uracil-containing DNA. Native DNA does not contain uracil so that the sample is not degraded by this procedure.

Source: Recombinant E. coli

Concentration and Size: 1U/μL

Applications：

qPCR and Digital PCR: Elimination of uracil from previous PCR products to prevent re-amplification in subsequent reactions, increasing assay sensitivity and specificity.

Next-Generation Sequencing (NGS): Critical component in NGS sample prep workflows to remove uracil bases from contaminating DNA molecules, improving sequence accuracy and reducing background noise.

Forensic Analysis: Enables more accurate and reliable detection of trace amounts of DNA in forensic samples by minimizing cross-contamination.

Methylation Detection: In conjunction with bisulfite conversion methods, UNG treatment ensures removal of unmethylated cytosines converted to uracil, facilitating methylation analysis.

Inhibitor Removal: Assists in purifying DNA by digesting degraded or damaged DNA containing uracil, thus enhancing the purity of target DNA sequences.

The Thermolabile Uracil-DNA Glycosylase Enzyme is an indispensable tool for molecular biologists and researchers engaged in sensitive nucleic acid-based experiments, offering superior control over DNA sample integrity and reducing unwanted artifacts that could compromise experimental outcomes.

Unit Definition

One unit is defined as the amount of Uracil-DNA Glycosylase (UNG), required to completely degrade 1μg of purified single-stranded uracil-containing DNA at 37°C in 60min.

Storage Buffer

20mM Tris-HCl (pH8.0), 0.1mM EDTA, 100mM KCl, 1mM DTT, 0.5%(v/v) Tween-20, 0.5%(v/v) NP-40, 50%(v/v) glycerol.

Quality Control

Purity>99% by SDS page

None contamination of Endonuclease, Nickase, Exonuclease and RNase.

Heat inactivation

50°C for 2 min

Reaction temperature

20~37°C for 10 min.

Amount of usage

0.1~1U of UNG enzymes per 50ul reaction.

Shipping and storage

Store at-20°C, ship with gel ice.