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| Product Name | Taq DNA Polymerase PCR Master Mix (2x) |
| Catalog No. | NATR-HMM-0113 |
| Description | A convenient 2× premixed solution containing thermostable Taq DNA polymerase, deoxynucleotide triphosphates (dNTPs), optimized reaction buffer with magnesium chloride, and PCR enhancers. The master mix reduces pipetting steps and minimizes contamination risk while providing robust amplification for a wide range of DNA templates. |
| Intended Use | Routine PCR amplification of DNA fragments up to 5 kb from genomic DNA, cDNA, plasmid DNA, and bacterial colonies for cloning, genotyping, mutation screening, and sequencing template preparation. |
| Principle / Technology | Thermostable Taq DNA polymerase catalyzes the template-directed incorporation of dNTPs into the growing DNA strand during thermal cycling. The enzyme is a recombinant form of Thermus aquaticus DNA polymerase expressed in E. coli. The 2× master mix format ensures consistent component concentrations across reactions. |
| Detection Method | Agarose gel electrophoresis with ethidium bromide or SYBR Safe staining; capillary electrophoresis; melt curve analysis; Sanger sequencing of purified products |
| Sample Type | Genomic DNA, plasmid DNA, cDNA, bacterial colonies (direct colony PCR), lambda phage DNA, purified PCR products |
| Performance Range / Specifications | Amplicon size range: 100 bp to 5 kb (standard), up to 10 kb with extended protocol; fidelity: approximately 1 error per 10^5 bases (error rate ~2.2 × 10^-5); amplification efficiency: >95% for targets up to 2 kb |
| Sensitivity / LOD | Reliably amplifies from as little as 1 pg of plasmid template or 10 pg of genomic DNA in a 50 μL reaction |
| Specificity | Minimal primer-dimer and non-specific product formation with optimized buffer system; no detectable E. coli genomic DNA contamination |
| Reaction Conditions / Protocol | Thaw and mix 2× master mix on ice, set up 50 μL reactions: 25 μL master mix, forward and reverse primers (0.1–1.0 μM each), template DNA, nuclease-free water to volume; typical thermal cycling: 95°C 3 min initial denaturation, 30–35 cycles of (95°C 30 sec, Tm–5°C 30 sec, 72°C 30–60 sec/kb), 72°C 5 min final extension |
| Components / Formulation | Taq DNA polymerase (0.05 U/μL final in 1× reaction), dNTPs (0.2 mM each final), Tris-HCl pH 8.7, KCl, (NH4)2SO4, MgCl2 (1.5 mM final), proprietary PCR enhancers and stabilizers, glycerol, bromophenol blue tracking dye (in some formulations) |
| Storage Conditions | –20°C for long-term storage; stable at 2–8°C for up to 3 months; stable at room temperature for setup; avoid repeated freeze-thaw cycles beyond 20 times |
| Shelf Life | 24 months at –20°C from date of manufacture |
| Package Specifications | 250 reactions (1.25 mL, based on 50 μL reaction volume), 1000 reactions (5 mL), 5000 reactions (25 mL) |
| Product Form | Liquid 2× master mix; direct use after thawing and mixing |
| Quality Control | Each lot functionally tested: PCR amplification of 0.5–3.0 kb fragments from human genomic DNA; absence of detectable endonuclease, exonuclease, and RNase activity; colony PCR performance verified with E. coli DH5α; negative control reactions show no amplification after 35 cycles |
| Key Features | 2× format reduces pipetting error and contamination; room-temperature reaction setup possible with antibody-mediated hot-start variant; compatible with high-throughput and automated liquid handling; green dye variant available for direct gel loading without loading buffer |
For research use only, not for clinical use.
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