DTI Uracil DNA Glycosylase(UNG), Heat-labile, (Glycerol Free) (2 U/ul) hydrolyzes N-glycosylic bonds between the deoxyribose sugars and the uracil bases in uracil-containing DNA leaving apyrimidinic sites in the DNA. UNG excises uracil from both single- and double-stranded dU-containing DNA but not from RNA. The glycerol free version makes the product suitable for lyophilisation.
| Brand | Catalogue Number | Pack Size |
|---|---|---|
|
DTI UNG 2 U/μl – Glycerol Free |
DT0105.20 |
20 U |
| Component | Pack Size |
|---|---|
|
DTI Uracil DNA Glycosylase (UNG), Heat-labile, (Glycerol Free) (2U/ul) |
10 µl |
|
Customized pack size |
Available upon request |
| Component | Concentration |
|---|---|
|
Tris-HCl, pH 8.0 |
20 mM (at 4℃) |
|
KCI |
100 mM |
|
DTT |
1 mM |
|
EDTA |
0.1 mM |
|
NP-40 |
0.5% (v/v) |
|
Tween 20 |
0.5% (v/v) |
Storage and Shipment Conditions: -80°C
Source: Produced in E. coli strain expressing a recombinant Xiphophorus maculatus UNG mutant.
Unit Definition: One unit of UNG is defined as the amount of enzyme required to digest 1 μg of uracil-containing dsDNA at 25℃ in 30 min.
Application: Further manufacturing and lyophilization processes for PCR, RT-PCR, and LAMP assays.
Inactivation by Heat: The enzyme is completely and irreversibly inactivated by heat incubating at 50℃ for 10 min.
Usage Protocol : Add suitable amount of enzyme based on the application. Generally, a range of 0.1 – 1 U/50μl of reaction volume is used.
UNG activation: Incubate for 10 min at 25℃.
UNG inactivation: Incubate for 2 min at 95℃.
Follow with the protocol of application.
Quality Control Data: Please see the certificate of analysis for each lot.
| Document | Download Link |
|---|---|
|
DTI UNG 2 U/μl – Glycerol Free – Protocol |
|
|
DTI UNG 2 U/μl – Glycerol Free – Certificate of Analysis |
Experimental Sample Performance Data: qPCR reactions were performed with two qPCR reaction mixes: One with UNG at a concentration of 2 U/μl and the other without UNG. Both these mixes were spiked with dU-containing amplicons, mimicking carryover contamination. The reaction mix without UNG generated a regular amplification curve, while the reaction mix containing UNG degraded the dU-containing amplicons, resulting in a significant reduction in amplification from the mimicking carryover contamination.
Figure 1: Comparison of qPCR reactions with and without UNG.
Conclusions: This experiment demonstrates the effectiveness of UNG in degrading dU-containing amplicons, significantly reducing the amplification from the mimicking carryover contamination in qPCR reactions.
Methods:
PCR cycling conditions for reactions with UNG:
1 cycle at 94°C for 30 sec, followed by 30 cycles of 94°C for 30 sec, 55°C for 30 sec, and 72°C for 60 sec.
PCR cycling conditions for reactions without UNG:
1 cycle at 94°C for 30 sec, followed by 30 cycles of 94°C for 30 sec, 55°C for 30 sec, and 72°C for 60 sec.