Aerosol Contamination in the Operational Environment as the Most Common Cause of False Positives in PCR Results: The Pioneering Discovery of Uracil DNA Glycosylase (UDG) by American Scientist Lindahl in E. coli and Bacillus subtilis, and the Establishment of a PCR Contamination Prevention System through the Use of UDG Enzyme with dUTP.
The commercially available UDG enzymes primarily consist of two types: the regular UDG enzymes derived from Escherichia coli and Bacillus subtilis, and the thermolabile UDG enzymes sourced from psychrophilic marine bacteria. The regular UDG enzymes exhibit greater heat resistance, retaining a small amount of uracil-DNA glycosylase activity even after treatment at 95°C for 10 minutes, which may lead to the degradation of target amplification products containing dU. Additionally, due to the use of engineered bacterial strains (such as E. coli) for the recombinant expression of molecular enzymes, there is a certain amount of residual host genomic DNA in these enzymes. Coupled with the influences of the manufacturing environment and human sources, molecular enzyme products are highly susceptible to DNA contamination. During the detection of pathogens, contaminating DNA may either mask low-abundance target nucleic acids or be detected alongside them, thereby affecting the interpretation of results.
Yeasen UCF.ME Uracil DNA Glycosylase (UDG/UNG), heat-labile
Description
The UCF.ME Uracil DNA Glycosylase (UDG/UNG), heat-labile, 1 U/μL (Cat#14466ES) derived from psychrophilic marine bacteria, it is an ultra-low residual heat-labile UDG enzyme known as UCF.ME. The use of this product can prevent the residual activity of conventional UDG enzymes after inactivation from degrading dU-containing amplification products at room temperature. It also prevents background bacteria present in molecular enzymes from causing false positives in PCR results. Therefore, the introduction of UCF.ME® ultra-low residual thermolabile UDG enzyme is crucial for accurately identifying infectious pathogens and assisting in the clinical diagnosis of infections and infectious diseases.
Product advantages
- Ultra-low residual of UCF.ME—E. coli genomic DNA residue <0.1 copies/ U;
- Low nuclease residue—No residual exonuclease, nicking enzyme, or RNase;
- Strong digestion capability—0.05 U/T can digest 105 copies/T dU-DNA products;
- Good thermolability—Complete inactivation under any condition of 50°C for 10 minutes, 55°C for 5 minutes, or 95°C for 5 minutes;
- Compatible with RT-qPCR reaction systems—No impact on the detection system even at high input levels (2U/20μL reaction system).
（1）Protein Purity≥95%
Figure 1. Protein purity (SDS-PAGE) detection results
（2）Low Residual Nucleases: No Residual Exonucleases, Nicking Enzymes, or RNases
Figure 2. Nuclease residue test results
（3）E. coli genomic DNA residue< 0.1 copies/ U，the residue levels were significantly lower than the conventional reagent
Figure 3. E.coli genomic DNA residue test results
Figure 4. A total of 0.05 U of UDG enzyme could completely digest 10^5 copies/T dU-DNA products.
Figure 5. After 50℃，10min；55℃，5min； 55℃，10min；95℃，5min heat inactivation treatment, 14466ES lost digestive ability.
Figure 6. RT-qPCR mix was prepared by using two priming probes of the ORF1ab and N genes and 14466ES. When 14466ES was injected into the reaction system at the amount of 2U/20μL, there was no inhibition of RT-qPCR reaction.
Product Recommendation — Ultra-Low Residual PCR Enzyme Raw Materials Series
|
Product Positioning |
Product Name |
Cat |
|
Contamination-Preventing heat-labile UDG |
UCF.ME Uracil DNA Glycosylase (UDG/UNG), heat-labile, 1 U/μL |
14466ES |
|
Hot start Taq DNA Polymerase |
Hieff UCF.ME Hotstart Sensitive Taq DNA Polymerase (5 U/μL) |
14314ES |
|
Reverse Transcriptase |
Hifair UCF.ME V Reverse Transcriptase (200 U/μL) |
14608ES |
|
Murine RNase Inhibitor |
14672ES |