UDG (uracil DNA glycosylase) can catalyze the hydrolysis of the N-glycosidic link between the uracil base and the sugar-phosphate backbone in ssDNA and dsDNA. It can easily control aerosol pollution and is suitable for common molecular biology systems such as PCR, qPCR, RT-qPCR and LAMP.
Component Number |
Components |
14455ES60 (100 U) |
14455ES76 (500 U) |
14455ES96 (10000 U) |
14455ES98 (25000 U) |
14455 |
Uracil DNA Glycosylase (UDG), 1 U/μL |
100 μL |
500 μL |
10 mL |
25 mL |
1. Remove aerosol pollution of dU-containing PCR products.
2. Remove uracil from single or double-stranded DNA.
One unit (U) is defined as the amount of enzyme that required to catalyze the hydrolysis of 1 μg dU-containing dsDNA in 30 minutes at 25°C.
95°C, 5~10 min.
The product is shipped with dry ice and can be stored at -25~-15℃ for 2 years.
Cycle step |
Temperature |
Time |
Cycles |
dU-containing template degradation |
25°C |
10 min |
1 |
UDG inactivation, template Pre-denaturation |
95°C |
5~10 min |
1 |
Denaturation |
95°C |
10 sec |
30-35 |
Annealing |
60°C |
20 sec |
|
Extension |
72°C |
30 sec/kb |
|
Final extension |
72°C |
5 min |
1 |
Note:The reaction time at 25°C can be adjusted within 5-10 min according to the experimental requirements.
Components |
Volume (μL) |
Final concentration |
10×PCR Buffer (Mg2+ Plus) |
5 |
1× |
25 mmol/L MgCl2 |
3 |
1.5 mmol/L |
dUTP (10 mmol/L) |
3 |
0.6 mmol/L |
dCTP/dGTP/dATP/dTTP (10 mmol/L each) |
1 |
0.2 mmol/L each |
Template DNA |
X |
- |
Primer 1 (10 μmol/L) |
2 |
0.4 μmol/L |
Primer 2 (10 μmol/L) |
2 |
0.4 μmol/L |
Taq DNA Polymerase (5 U/μL) |
0.5 |
0.05 U/μL |
Uracil DNA Glycosylase (UDG), 1 U/μL |
1 |
1 U/50 μL |
ddH2O |
Up to 50 |
- |
Note:
According to the experimental requirements, the final concentration of dUTP can be adjusted between 0.2-0.6 mmol/L, and 0.2 mmol/L dTTP can be added selectively.
2.Amplification procedure
Cycle step |
Temperature |
Time |
Cycles |
dU-containing template degradation |
25°C |
10 min |
1 |
UDG inactivation, template Pre-denaturation |
95°C |
5~10 min |
1 |
Denaturation |
95°C |
10 sec |
30-35 |
Annealing |
60°C |
20 sec |
|
Extension |
72°C |
30 sec/kb |
|
Final extension |
72°C |
5 min |
1 |
Note:The reaction time at 25°C can be adjusted within 5-10 min according to the experimental requirements.
1. UDG is active in most PCR reaction buffers.
2. Enzymes should be stored in an ice box or on an ice bath when used, and should be stored at -20°C immediately after use.
3. For your safety and health, please wear lab coats and disposable gloves for operation.
4. This product is for research use ONLY!
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