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| Product Name | Hot-Start Taq DNA Polymerase 2× Master Mix |
| Catalog No. | NATR-HMM-0114 |
| Description | A 2× concentrated PCR master mix featuring antibody-mediated hot-start Taq DNA polymerase that remains inactive at room temperature and is activated during the initial denaturation step. This hot-start mechanism effectively suppresses primer-dimer formation and non-specific amplification during reaction setup. |
| Intended Use | High-specificity PCR amplification for applications requiring minimal background, including mutation detection, allele-specific PCR, low-copy template amplification, multiplex PCR, and clinical genotyping assays. |
| Principle / Technology | Taq DNA polymerase is complexed with monoclonal antibodies that reversibly inhibit enzymatic activity below 70°C. During the initial denaturation at 94–95°C, antibodies are irreversibly denatured and released, restoring full polymerase activity. This prevents extension of misprimed templates and primer-dimers formed during reaction setup and the initial temperature ramp. |
| Detection Method | Agarose gel electrophoresis; high-resolution melt analysis; Sanger sequencing; fragment analysis by capillary electrophoresis; probe-based or intercalating dye detection in real-time PCR |
| Sample Type | Genomic DNA from blood, tissue, FFPE, and forensic samples; cDNA; plasmid DNA; low-copy-number targets; crude lysates from bacterial colonies and cell cultures |
| Performance Range / Specifications | Amplicon length: 100 bp to 4 kb; amplification efficiency: 90–110% for targets 100–500 bp; activation time: 2–5 minutes at 95°C; error rate: ~2.2 × 10^-5 errors per base per duplication |
| Sensitivity / LOD | Detects single-copy genomic targets in human genomic DNA (as low as 3 pg); reliably amplifies from <10 copies of plasmid template per 50 μL reaction |
| Specificity | Antibody-mediated hot-start provides >100-fold reduction in non-specific amplification products compared to standard Taq; no detectable amplification in no-template controls after 40 cycles; does not interfere with downstream sequencing and cloning |
| Reaction Conditions / Protocol | Set up reactions on ice or at room temperature: 25 μL master mix, primers at 0.2–1.0 μM each, template DNA, nuclease-free water to 50 μL total; recommended cycling: 95°C 2–5 min (antibody denaturation), 30–40 cycles of (95°C 15–30 sec, 55–65°C 15–30 sec, 72°C 30–60 sec/kb), 72°C 5 min final extension; hold at 4°C |
| Components / Formulation | Hot-start Taq DNA polymerase (~0.05 U/μL final), dNTPs (0.2 mM each final), KCl and (NH4)2SO4, MgCl2 (1.5–2.0 mM final), stabilizers, antibody complex, Tris-HCl pH 8.5, PCR enhancers |
| Storage Conditions | –20°C for long-term storage; stable at 2–8°C for up to 6 months; room-temperature setup stable for at least 72 hours without loss of hot-start specificity |
| Shelf Life | 24 months at –20°C from date of manufacture |
| Package Specifications | 250 reactions (1.25 mL), 1000 reactions (5 mL), and bulk packaging options available |
| Product Form | Liquid 2× master mix, ready to use |
| Quality Control | Each lot tested: single-copy gene amplification from 10 ng human genomic DNA; no non-specific bands in no-template control; DNase/RNase activity absent; PCR efficiency 90–110% across 5-log dynamic range by qPCR; functional with GC-rich (>70%) templates |
| Key Features | Room-temperature stable reaction assembly; proven antibody hot-start provides >100-fold specificity improvement; long shelf life at –20°C; compatible with direct PCR from colonies and crude lysates; suitable for high-GC-content templates with separate GC-rich buffer |
For research use only, not for clinical use.
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