Uracil-DNA Glycosylaes (UDG)

Product Description

Thermolabile Uracil DNA Glycosylase (Thermolabile UDG) was derived from E.coli recombination, cloning, and expression UDG gene from a psychrophilic marine bacterium. Thermolabile UDG catalyzes the hydrolysis of the N-glycosylic bond between uracil and glycophosphate skeleton, leaving an apyrimidinic site in uracil-containing single or double-stranded DNA. This product is sensitive to high temperatures and can be irreversibly inactivated above 55℃. This product is suitable for PCR, qPCR, RT-PCR, and RT-qPCR systems.

Storage Temperature

Store at -20 ℃, valid for 1 year.

Storage Buffer

20 mM Tris-HCl (pH 8.0, 25℃), 0.1 mM EDTA, 100 mM KCl, 50% Glycerol (v/v).

Unit Definition

One unit is defined as the amount of enzyme that catalyzes the release of 60 pmol of uracil per minute from double-stranded, uracil-containing DNA. The activity is determined by the amount of [3H]-uracil released from a 50 μL reaction system containing 0.2 μg DNA (104-105 cpm/μg) at 37℃ in 30 minutes.

Molecular Weight of Protein

The molecular weight of Thermolabile UDG is 25 kDa.

Quality Control Endonuclease Activity:

Incubation of a 50 µL reaction containing a minimum of 1 unit of Thermolabile UDG with 1 μg λDNA at 37 ℃ for 16 hours yielded no degradation as determined by agarose gel electrophoresis.

Exonuclease Activity:

Incubation of a 50 µL reaction containing a minimum of 1 unit of Thermolabile UDG with 1 μg λ-Hind III digest DNA at 37℃ for 16 hours yielded no degradation as determined by agarose gel electrophoresis.

Nicking Activity:

Incubation of a 50 µL reaction containing a minimum of 1 unit of Thermolabile UDG with 1 μg pBR322 at 37 ℃ for 16 hours resulted in <10% conversion to the nicked form as determined by agarose gel electrophoresis.

RNase Activity:

Incubation of a 50 µL reaction containing a minimum of 1 unit of Thermolabile UDG with 1.6 μg MS2 RNA for 4 h at 37 ℃ yielded no degradation as determined by agarose gel electrophoresis.

E.coli DNA: A minimum of 1 unit of Thermolabile UDG was screened for the presence of E. coli genomic DNA using TaqMan qPCR with primers specific for the E. coli 16s rRNA locus. Results were quantified using a standard curve generated from purified E. coli genomic DNA. The measured level of E. coli genomic DNA contamination was ≤1 E. coli genome.

Product Application

This product can be widely used in any experiment, such as: 1. Remove Carry-over Contamination in PCR. 2. Remove Uracil-base from single – or doublestranded DNA. 3. Glycosylase-mediated single nucleotide polymorphism detection (GMPD).

Usage Note

1. Thermolabile UDG is active in most PCR or RT-PCR systems, but for personal systems, it is recommended to test whether it is compatible with the system used for the first time.
2. For your safety and health, please wear a lab coat and disposable gloves to operate.