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| Product Name | Standard Taq DNA Polymerase |
| Catalog No. | IEC-HMM-0048 |
| Description | A recombinant form of the thermostable DNA polymerase originally isolated from Thermus aquaticus. The enzyme catalyzes template-directed incorporation of deoxynucleotides at the 3'-hydroxyl terminus of a primer annealed to a complementary DNA template. Exhibits 5'-3' polymerase and 5'-3' exonuclease activities but lacks 3'-5' proofreading function. |
| Intended Use | For conventional endpoint PCR, colony PCR, genotyping, microsatellite analysis, and general-purpose DNA amplification from a variety of template sources. |
| Principle / Technology | Template-dependent DNA synthesis via nucleotidyl transfer from deoxynucleoside triphosphates to the 3'-OH terminus of a primer-template duplex, proceeding in the 5' to 3' direction along the template strand. |
| Detection Method | Activity quantified by incorporation of [³H]-dTTP into acid-insoluble precipitate at 74 °C using nicked calf thymus DNA as substrate. |
| Sample Type | Genomic DNA (human, animal, plant, microbial), plasmid DNA, phage DNA, cDNA, and bacterial colonies. |
| Performance Range / Specifications | Specific activity >= 20,000 U/mg. Extension rate: 1-2 kb/min at 72 °C. Amplicon range: 100 bp to 6 kb optimal. Generates 3'-A overhang suitable for TA cloning. |
| Sensitivity / LOD | Reliably amplifies single-copy genomic targets from 50-100 pg template DNA in a 25 uL reaction after 30-35 cycles. |
| Specificity | Standard polymerization fidelity; error rate ~2.2 x 10⁻⁵ errors per base per duplication. 5'-3' exonuclease activity enables TaqMan probe hydrolysis. |
| Reaction Conditions / Protocol | Recommended cycling: 95 °C initial denaturation 3 min; 25-35 cycles of (95 °C 30 s, TM-5 °C 30 s, 72 °C 1 min/kb); final extension 72 °C 5 min. |
| Components / Formulation | Taq DNA Polymerase (5 U/uL) in 20 mM Tris-HCl (pH 8.0), 100 mM KCl, 0.1 mM EDTA, 1 mM DTT, 50% glycerol, 0.5% Tween-20, 0.5% NP-40. Supplied with 10x PCR Buffer and 50 mM MgCl₂. |
| Storage Conditions | Long-term storage at -20 °C to -30 °C. Short-term storage (up to 2 weeks) at 4 °C acceptable without significant activity loss. |
| Shelf Life | 36 months at -20 °C from date of manufacture. |
| Package Specifications | 200 U, 500 U, 1,000 U, 5,000 U, and 10,000 U configurations. Each includes proportional volumes of 10x buffer and MgCl₂ solution. |
| Product Form | Clear, colorless to faint yellow liquid with glycerol as stabilizer. |
| Quality Control | Each lot verified for: specific activity (>= 20,000 U/mg); absence of contaminating DNase, RNase, and non-specific endonuclease; functional PCR performance across multiple template types and amplicon sizes. |
| Key Features | Robust performance across diverse template types; produces 3'-A overhangs compatible with TA cloning vectors; extensive published validation; economical for routine PCR applications. |
| Purity | >= 95% pure by SDS-PAGE densitometry. No detectable contaminating nuclease activities. |
| Concentration | 5 U/uL standard formulation. 10 U/uL high-concentration variant available. |
| Activity / Unit Definition | 1 unit = 10 nmol dNTP incorporated in 30 min at 74 °C. |
| Molecular Weight | ~94 kDa (monomer protein subunit). |
| Source / Origin | Recombinant expression in E. coli from Thermus aquaticus DNA polymerase I gene. No animal-derived components in production process. |
| pH Range / Optimal pH | Optimal pH 8.0-9.0 (Tris-based buffer). Operable range pH 7.5-9.5. |
| Shipping Conditions | Shipped with cold packs to maintain below 10 °C. Arrival at ambient temperature (<72 h) does not affect activity if promptly stored at -20 °C. |
| Expiration Date / Stability | >90% activity retention after 36 months at -20 °C. Degradation accelerates above 0 °C; avoid prolonged room temperature exposure. |
| Regulatory / Compliance | ISO 9001 and ISO 13485 certified manufacturing. Suitable as IVD raw material; regulatory documentation for product registration available on request. |
| Compatibility | Compatible with all common thermal cycler instruments. Works with DMSO (<= 10%), glycerol (<= 15%), betaine (<= 1.5 M). Requires Mg²⁺ optimization for each primer set. |
| Recommended Buffer System | Standard 10x buffer: 200 mM Tris-HCl (pH 8.8), 100 mM KCl, 100 mM (NH₄)₂SO₄, 20 mM MgSO₄, 1% Triton X-100. Detergent-free buffer variant available for mass spectrometry applications. |
| Application Notes / Precautions | Set up reactions on ice. For colony PCR, pick a small colony and resuspend directly in the reaction mix. Adjust Mg²⁺ between 1.5-4.0 mM final for optimal results. GC-rich templates benefit from 3-10% DMSO or 1 M betaine. |
| Batch-to-Batch Consistency | Inter-lot activity variation <= ±10%. Comparable PCR performance across production batches validated with standardized template/primer panels. |
For research use only, not for clinical use.
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