DTI UNG 2 U/μl - Glycerol Free
DTI UNG 2 U/μl - Glycerol Free
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.
SKU:DT0105.20
DTI UNG 2 U/μl - Glycerol Free
| Brand | Catalogue Number | Pack Size |
|---|---|---|
| DTI UNG 2 U/μl - Glycerol Free | DT0105.20 | 20 U |
Components
| Component | Details |
|---|---|
| DTI Uracil DNA Glycosylase (UNG), Heat-labile, (Glycerol Free) (2U/ul) | 10 µl |
| Customized pack size | Available upon request |
Storage Buffer
| 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.
DTI UNG 2 U/μl - Glycerol Free - Protocol and Downloads
| Document | Download Link |
|---|---|
| DTI UNG 2 U/μl - Glycerol Free - Protocol | Download User Manual |
| DTI UNG 2 U/μl - Glycerol Free - Certificate of Analysis | Download CoA - DT0105.20 |
| DTI UNG 2 U/μl - Glycerol Free - Promotional Document | Download Flyer |
Results
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.