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| Product Name | dUTP Mix (10 mM, PCR Carryover Prevention Grade) |
| Catalog No. | IEC-HMM-0060 |
| Description | A nucleotide mixture containing dATP, dCTP, dGTP (10 mM each) and dUTP (20 mM) formulated to replace standard dTTP in PCR for carryover contamination prevention. When used with uracil-DNA glycosylase (UDG), any contaminating products from previous amplifications containing uracil are enzymatically degraded before new amplification begins. |
| Intended Use | For PCR carryover contamination control in combination with UDG. Essential component of the dUTP/UDG carryover prevention system for diagnostic PCR, high-throughput screening, and any laboratory performing repeated amplification of the same target sequences. |
| Principle / Technology | dUTP replaces dTTP during PCR amplification, generating uracil-containing amplicons. In subsequent reactions, UDG pre-treatment specifically degrades any uracil-containing carryover DNA from prior amplifications while leaving natural thymine-containing template DNA intact and amplifiable. |
| Detection Method | HPLC purity analysis and functional carryover prevention testing with standardized UDG protocol. |
| Sample Type | Used in PCR master mixes for carryover-sensitive amplification workflows. |
| Performance Range / Specifications | dATP, dCTP, dGTP: 10 mM each. dUTP: 20 mM. Total available incorporation capacity matches DNA polymerase requirements for standard and long-range PCR. dUTP at 20 mM enables complete replacement of dTTP at recommended working concentrations. |
| Sensitivity / LOD | Supports amplification sensitivity equivalent to standard dNTP/dTTP mixtures when used at 200 uM dUTP final concentration. |
| Specificity | DNA polymerases incorporate dUTP with efficiency comparable to dTTP. UDG specifically recognizes and cleaves uracil bases, leaving thymine-containing natural templates intact. |
| Reaction Conditions / Protocol | Replace standard dNTP mix with dUTP Mix at same working concentration. Use 200 uM each dATP, dCTP, dGTP and 400 uM dUTP final. Combine with 0.5-1 U UDG per reaction and pre-incubate at 37 °C 10 min. |
| Components / Formulation | 10 mM dATP, 10 mM dCTP, 10 mM dGTP, 20 mM dUTP in ultrapure nuclease-free water, pH 7.0-7.5. |
| Storage Conditions | Store at -20 °C to -30 °C protected from light. Aliquot for single use. |
| Shelf Life | 24 months at -20 °C. |
| Package Specifications | 0.25 mL, 1 mL, 5 mL, 25 mL configurations. Compatible UDG enzyme available separately or as a combined kit. |
| Product Form | Clear, colorless aqueous solution. pH neutral. |
| Quality Control | Each lot tested by: HPLC purity (>= 99% per nucleotide); UDG substrate functionality; PCR performance compared to standard dNTP mix; carryover degradation efficiency (>99% digestion of dU-containing 500 bp amplicon). |
| Key Features | Enables closed-tube carryover prevention when combined with UDG; dUTP concentration optimized for complete dTTP replacement; compatible with most DNA polymerases; essential for high-sensitivity diagnostic PCR. |
| Purity | >= 99% per individual dNTP by HPLC. dUTP purity independently verified at >= 99%. |
| Concentration | dATP, dCTP, dGTP: 10 mM each. dUTP: 20 mM. |
| Activity / Unit Definition | DNA polymerase utilization efficiency: dUTP incorporated at approximately 85-95% of dTTP efficiency depending on polymerase type. Compensated by higher dUTP concentration (20 mM vs 10 mM). |
| Molecular Weight | dUTP (trisodium): ~550.1 g/mol. |
| Source / Origin | Synthesized via enzymatic or chemical phosphorylation of purified deoxynucleoside precursors. Produced in ISO-certified nucleotide manufacturing facility. |
| pH Range / Optimal pH | Solution pH 7.0-7.5. Neutral pH minimizes dUTP deamination during storage. |
| Shipping Conditions | Shipped with cold packs to maintain below 10 °C. Transport at ambient temperature for <1 week acceptable. |
| Expiration Date / Stability | 24 months at -20 °C. dUTP is slightly more labile than dTTP; avoid pH extremes and elevated temperatures. Aliquot to minimize freeze-thaw exposure. |
| Regulatory / Compliance | ISO 9001 and ISO 13485 certified. Suitable for IVD kit incorporation with carryover prevention feature. |
| Compatibility | Compatible with Taq and most Taq-derived DNA polymerases. Not recommended for archaeal family B polymerases (Pfu, Vent, Deep Vent) which do not efficiently incorporate dUTP. Compatible with antibody-mediated and chemical hot-start polymerases. |
| Recommended Buffer System | Supplied in nuclease-free water. Dilute into PCR reaction buffer at point of use. |
| Application Notes / Precautions | dUTP-containing PCR products may show altered electrophoretic mobility compared to dTTP products. For diagnostic applications, validate the carryover prevention protocol with the specific primer sets and thermal cycling conditions in use. dUTP-based amplicons are not suitable for downstream restriction digestion by all enzymes; verify compatibility. |
| Batch-to-Batch Consistency | HPLC purity and functional carryover prevention performance verified for every production lot. Lot-to-lot PCR Ct values within +/- 0.5 cycles of reference. |
For research use only, not for clinical use.
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