UCF.ME® Uracil DNA Glycosylase(UDG/UNG), heat-labile, 1 U/μL

Product description

Heat-Labile UDG (uracil DNA glycosylase) catalyzes the release of free uracil from uracil-containing DNA by hydrolyzing the N-glycoside bond between uracil bases and sugar phosphate skeletons. Compared with conventional UDG enzyme, heat-Labile UDG can avoid the degradation of dU-containing amplification products caused by residual activity of the inactivated UDG. This product is sensitive to temperature, and can be completely inactivated by incubation at 55℃ for 5 min or 50℃ for 10 min. In addition, this product has been processed by the UCF.ME^TM ultra-low residue process, and its residual DNA contamination of E. coli  is extremely low, which is suitable for the detection of pathogenic microorganisms and other fields.

Specifications

Cat. No.	14466ES60 / 14466ES76 / 14466ES96
Size	100 U / 500 U / 10,000 U
Heat Inactivation	55℃ for 5 min; 50℃ for 10 min

Components

Name	14466ES60	14466ES76	14466ES96
UCF.METM Uracil DNA Glycosylase (UDG/UNG), heat-labile, 1 U/μL	100 μL	500 μL	10 mL

Storage

The product should be stored at -25~-15°C for 2 years.

Instructions

1. Reaction Setup

Components	Volume (μL)	Final Concentration
10×PCR Buffer (Mg2+ Plus)	5	1×
25 mM MgCl2	3	1.5 mM
dUTP (10 mM)	3*	0.6 mM
dCTP / dGTP/ dATP/ dTTP (10 mM each)	1	0.2 mM each
Template DNA	Optional	-
Primer 1 (10 μM)	2	0.4 μM
Primer 2 (10 μM)	2	0.4 μM
Taq DNA Polymerase (5 U/μL)	0.5	0.05 U/μL
UCF.METM Uracil DNA Glycosylase (UDG/UNG), heat-labile, 1 U/μL	1	1 U/50 μL
ddH2O	Up to 50	-

*According to the demands of the experiment, the final concentration of dUTP can be adjusted between 0.2~0.6 mM, and 0.2 mM dTTP can be added selectively. 

2. Amplification Procedure

Stage	Temperature	Time	Cycles
dU-containing template degradation	25℃	10 min*	1
UDG inactivation and pre-denaturation of template	94℃	2 min	1
Denaturation	95℃	10 sec	30~35
Annealing	60℃	20 sec
Extension	72℃	30 sec/kb
Final extension	72℃	5 min	1

*The reaction time at 25℃ can be adjusted within 5~10 min according to the experimental requirements.

 Notes

1. Heat-labile UDG is active in most PCR reaction buffers.

2. The enzyme should be stored in the ice box or ice bath when used, and should be stored at -25~-15°C immediately after use.

3. For your safety and health, please wear lab coats and disposable gloves for operation.

4. This product is scientific research purposes only.

Ver.EN20230922