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Uracil DNA Glycosylase (UDG/UNG), heat-labile

Product SKU

Specifications

Price

10303ES60

100 U

$70.25

10303ES76

500 U

$301.85

10303ES96

10000 U

$4825.77

10303ES98

100000 U

$39318.73

Introduction

Heat-Labile UDG (uracil DNA glycosylase) excises uracil from dU-containing DNA by catalysing and hydrolysing uracil bases and N-glycoside bonds of the sugar phosphate skeletons, releasing free uracil. Compared with ordinary UDG enzyme, Heat-Labile UDG can avoid the degradation of dU-containing amplified products at room temperature by residual activity of ordinary UDG. The product works at room temperature and is sensitive to temperature and easy to be inactivated.

Application

Remove aerosol pollution of dU-containing PCR products.

Remove uracil bases of single or double-stranded DNA.

Package Information

Component Number

Component

10303ES60 (100 U)

10303ES76 (500 U)

10303ES92 (10000 U)

10303-A

Uracil DNA Glycosylase (UDG), Heat-Labile (1 U/μL)

100 μL

500 μL

10 mL

Unit Definition

One unit (U) is the amount of enzyme that required to liberate 1 μg uracil from dU-containing DNA in 30 minutes at 25°C.

Inactivation mode

50℃, 10 min.

Storage

Dry ice transportation. Store at -20℃.

Note

For your safety and health, please wear experimental clothes and disposable gloves.

Heat-Labile UDG was found to be active in most PCR buffers.

Quality Control

Endonuclease residue detection: DNA electrophoresis bands do not change when 10 U of this enzyme and 0.6 μg Hind III digested λDNA- are incubated at 37°C for 4 hours.

RNase residue detection: RNA electrophoresis bands do not change when 10 U of this enzyme and 1 μg of total RNA of HeLa cell are incubated at 37°C for 1 hour.

E.coil DNA residue detection: E.coil Genome residue of 200 U of this product should be less than 10 copies in TaqMan qPCR detection specified with 16s rDNA .

Enzyme inactivation test: DNA electrophoresis bands do not change when the product was treated at 50 ℃ for 10 min and incubated with 1 μg of dU-containing dsDNA template for 30 min.

Application example

Recommended reaction mixture for PCR

Components

Volume(μL)

Final concentration

10×PCR Buffer (Mg2+ Plus)

5

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

Heat-Labile UDG (1 U/μL)

1

0.05 U/μL

ddH2O

Up to 501

-

Note

According to the experimental requirements, the final concentration of dUTP can be adjusted between 0.2-0.6 mM. 0.2 mMdTTP can be added selectively

PCR conditions

Temperature

Time

Cycles

Stage

25℃

10 min

1

dU-containing template degradation

94℃

2 min

1

UDG inactivation, template Pre-denaturation

95℃

10 sec

-

Denaturation

60℃

20 sec

30-35

Annealing

72℃

30 sec/kb

-

Extension

72℃

5 min

1

Final extension

Note

The reaction time at 25 ℃ can be adjusted within 5-10 min according to the experimental needs.

15221439726

15221439726