BioReady Uracil DNA Glycosylase (UDG)
BioReady Uracil-DNA Glycosylase (UDG) is derived from recombinant cloning and expression in Escherichia coli. It effectively hydrolyzes uracil (U) in single-stranded or double-stranded DNA longer than 6 base pairs, with no activity on RNA. Its primary application is to prevent contamination of PCR amplification products. It works by replacing dTTP with dUTP in the PCR reaction, creating PCR amplification products containing dU bases. UDG then cleaves the dU bases in these amplification products, creating abasic sites, preventing the DNA strand from serving as a template for subsequent PCR, thus achieving the goal of preventing PCR product contamination. Additionally, this UDG becomes inactive in the subsequent 95°C pre-denaturation step, without affecting the amplification of the template.
Product Description
BioReady Uracil DNA Glycosylase catalyzes the release of free uracil from longer then 6bp dU-containing single-stranded or double-stranded DNA whereas RNA and normal dT-containing DNA. UDG excises uracil form dU-containing PCR product when dTTP replace with dUTP was incorporated into DNA. The resulting abasic sites are blocked from replication by PCR. It mainly used to prevent contamination of PCR amplification products. This enzyme can be completely and irreversibly inactivated after at temperatures above 95°C.
Application Field
Polymerase Chain Reaction (PCR)
Site-Directed Mutagenesis
Cloning
Sequencing
Genotyping
Gene Expression Analysis
Application Cases
1. The PCR reaction with UDG and without UDG, the amplification of templates containing uracil (U) was evaluated after digestion at 37°C for 2 minutes.
Results:
The results demonstrate that BioReady Uracil-DNA Glycosylase has a significant activity in degrading PCR products containing uracil (U). When using PCR products containing U as templates, the average amplification Ct in qPCR reactions without UDG is 17.83, whereas in qPCR reactions with UDG added, the average amplification Ct is 34.73. The difference is 16.9, indicating a more than 100,000-fold reduction in contaminated PCR products.
Storage Conditions
1. The kit needs to be shipped refrigerated.
2. Kit storage: Store in a -20°C refrigerator away from light, and the repeated freezing and
thawing should not more than 7 times.
3. Validity period: 12 months, please use within the validity period.
Ordering Information
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