BioReady Heat-labile UDG

Name: BioReady Heat-labile UDG
Brand: Bioer Technology

BioReady Thermosensitive 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 thermosensitive UDG becomes inactive in the subsequent 50°C reverse transcription or 95°C pre-denaturation steps without affecting the amplification of the template.

Product Description

BioReady Heat-labile UDG（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 is sensitive to heat. It can be completely and irreversibly inactivated after 10 min at temperatures above 50°C.

Product Features

Thermosensitive, heat inactivated at 50°C for 10 minutes.

Wide applicability, compatible with one-step RT-PCR systems.

Application Field

Polymerase Chain Reaction (PCR)

Site-Directed Mutagenesis

Cloning

Sequencing

Genotyping

Gene Expression Analysis

Application Cases

Experiments were conducted using one-step RT-qPCR reagents with and without BioReady Thermosensitive Uracil-DNA Glycosylase (BSA51) to evaluate the amplification of templates containing uracil (U) after digestion

at 25°C for 2 minutes.

Results:The results demonstrate that BioReady Thermosensitive Uracil-DNA Glycosylase (UDG) effectively degrades PCR products containing uracil (U). In the presence of PCR products containing U, the one-step RT-qPCR without UDG had an average amplification Ct of 29.66, while the one-step RT-qPCR with the inclusion of thermosensitive UDG showed no amplification at all.

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.