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| Product Name | Uracil DNA Glycosylase (UDG), Glycerol-Free |
| Catalog No. | IVDRM-0261 |
| Description | Uracil DNA Glycosylase (UDG) catalyzes the hydrolysis of the N-glycosidic bond between the uracil base and the sugar-phosphate backbone in uracil-containing DNA strands, releasing free uracil. It is used to eliminate aerosol carry-over contamination from dU-containing PCR products and to remove uracil bases from single- or double-stranded DNA. This glycerol-free formulation is well suited for lyophilized reagent development and IVD assay manufacturing. |
| Intended Use | Elimination of carry-over (aerosol) contamination from dU-containing PCR products Removal of uracil bases from single-stranded or double-stranded DNA |
| Principle / Technology | UDG excises uracil bases from DNA by hydrolyzing the N-glycosidic bond linking uracil to the deoxyribose-phosphate backbone, generating abasic (AP) sites that render dU-containing DNA non-amplifiable, thereby preventing carry-over contamination in dUTP-based PCR workflows. |
| Sample Type | PCR reaction mixtures containing dUTP; single- or double-stranded uracil-containing DNA |
| Reaction Conditions / Protocol | Example 50 uL PCR setup: 10x PCR Buffer (Mg2+ Plus): 5 uL 25 mmol/L MgCl2: 3 uL dUTP: 0.6 mmol/L dCTP / dGTP / dATP: 0.2 mmol/L each Template DNA: optional Primer 1 (10 umol/L): 2 uL Primer 2 (10 umol/L): 2 uL Taq DNA Polymerase (5 U/uL): 0.5 uL UDG (1 U/uL): 1 uL ddH2O: to 50 uL Cycling program: 25 °C, 10 min, 1 cycle - degrade U-containing templates 94 °C, 2 min, 1 cycle - UDG inactivation and template pre-denaturation 95 °C, 10 sec; 60 °C, 20 sec; 72 °C, 30 sec/kb, 30-35 cycles - denaturation, annealing, extension 72 °C, 5 min, 1 cycle - final extension |
| Storage Conditions | Store at 2-8 °C |
| Shelf Life | 6 months at 2-8 °C |
| Package Specifications | 100 U; 500 U |
| Product Form | Liquid, glycerol-free |
| Activity / Unit Definition | One unit (U) is defined as the amount of enzyme required to completely degrade 1 ug of uracil-containing dsDNA within 30 min at 25 °C. |
| Recommended Buffer System | Standard PCR buffer with Mg2+ (e.g., 10x PCR Buffer (Mg2+ Plus) plus additional 25 mmol/L MgCl2) |
| Application Notes / Precautions | The final dUTP concentration can be adjusted between 0.2-0.6 mmol/L as needed; 0.2 mmol/L dTTP may optionally be incorporated The 25 °C UDG treatment time can be adjusted within 5-10 min according to experimental needs This product is a conventional UDG and is fully inactivated only by heating at 95 °C for 5-10 min; it is not a heat-labile enzyme |
| Heat Inactivation | 95 °C for 5-10 min (this is a conventional, non-heat-labile UDG) |
For research use only, not for clinical use.
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