Heat Labile UDG
For research use only
R5001 (500 U)
Heat Labile UDG (1 U/μl)
This reagent should be kept at -30~15°C.
Heat Labile UDG is purified from the expression product of an recombinant E. coli strain containing the UDG gene cloned from cold-adaptive marine bacteria (psychrophilic Marine bacterium). UDG (Uracil-DNA Glycosylase) catalyzes the hydrolysis of uracil bases of the single or double stranded DNA containing dU and the N-glycoside bonds of the glycosphosphoric acid skeleton to release free uracil. The resulting base-free site is easily fractured by hydrolysis. This product is sensitive to high temperature, and above 50°C can make the enzyme irreversible inactivation, suitable for PCR, qPCR, RT-PCR, RT-qPCR system.
20 mM Tris-HCl, pH 8.0 at 25°C
0.1 mM EDTA
100 mM KCl
1 mM DTT
Glycerol 50% (v/v)
NP - 40 0.5% (v/v)
0.5% Tween - 20 (v/v)
In 70 mM Tris-HCl, pH 7.5, 10 mM NaCl, 1 mM EDTA, 100 g/ml BSA reaction solution, the amount of enzyme required to release 1 nmol uracil from DNA containing dU within 1 h at 37°C was defined as 1 unit of activity (U).
Exonuclease residue detection: 20 U of this product and 0.3 μg of λ-Hind III were incubated at 37°C for 16 h, and the DNA bands did not change after electrophoresis.
Detection of endonuclease residue: 20 U of this product and 0.3 μg of Supercoiled pBR322 DNA were incubated at 37°C for 4 h, and the DNA bands did not change after electrophoresis.
RNase residue detection: the product of 20 U and 1 μg of HeLa cell RNA were incubated at 37°C for 30 min, and the electrophoresis band of RNA was unchanged by agarose gel electrophoresis.
E.coli DNA residue detection: the nucleic acid residue in 20 U of this product was detected by E. coli gDNA-specific TaqMan qPCR, and the residue of E. coli genome was less than 10 copies.
1. Prepare the PCR reaction system as follows:
10 × Taq Buffer (Mg2+ plus)
0.2 mM each
Primer 1 (10 μM)
Primer 2 (10 μM)
Taq DNA Polymerase (5 U/μl)
Heat Labile UDG (1 U/μl)b
to 50 μl
a. According to the needs of the experiment, the final dUTP concentration can be adjusted between 0.2-0.6 mM.
b. According to the needs of the experiment, the amount of 50 μl reaction system is generally 0.1-1 U.
* According to the experimental needs, the final concentration of MgCl2 can be adjusted between 2-3 mM.
2. Reaction procedure
degradation of U - containing templates
UDG inactivation, template denaturation
* The PCR reaction procedure can be adjusted according to the experimental needs.