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| Product Name | Veritas Long-Range High-Fidelity PCR Master Mix (No Dye) |
| Catalog No. | RESK-0033 |
| Description | The Veritas Long-Range High-Fidelity PCR Master Mix (No Dye) is a ready-to-use 2x premixed solution containing a high-fidelity DNA polymerase, dNTPs, and a buffer system carefully optimized for long-range amplification, with an extension speed of 10 sec/kb. Its fidelity is 83x that of Taq DNA polymerase and 9x that of conventional Pfu DNA polymerase, making it well suited for the amplification of complex templates. Amplification products are blunt-ended. The master mix is fast and easy to use, with high sensitivity, strong specificity, and excellent stability, and it covers templates with 20-80% GC content. It can amplify whole blood samples without nucleic acid extraction - including blood preserved with EDTA, citrate, or heparin, as well as dried blood spot cards - for faster and more flexible workflows. A proprietary protective agent keeps the premix active even after repeated freeze-thaw cycles. All reagent components undergo strict quality control to ensure superior performance and batch-to-batch consistency. |
| Intended Use | Long-range, high-fidelity PCR amplification of DNA templates (genomic DNA, cDNA, plasmids, whole blood, and crude samples), especially complex templates; for research use. |
| Principle / Technology | A high-fidelity DNA polymerase with proofreading (3' to 5' exonuclease) activity combined with a buffer system optimized for long-range amplification enables accurate, rapid synthesis of long amplicons (83x Taq fidelity, 9x Pfu fidelity). Products are blunt-ended. This dye-free formulation leaves products free of tracking dye for downstream applications. |
| Sample Type | Genomic DNA (human, animal, plant), cDNA (including uracil-containing cDNA with residual RNA), plasmid DNA, lambda DNA, bacterial culture, and whole blood without extraction (EDTA-, citrate-, or heparin-preserved blood; dried blood spot cards). |
| Performance Range / Specifications | Amplifies DNA fragments up to 40 kb and cDNA fragments of 1-14 kb Extension speed: 5-10 sec/kb (10 sec/kb standard; 5 sec/kb for fragments below 10 kb) Fidelity: 83x that of Taq DNA polymerase and 9x that of conventional Pfu DNA polymerase Compatible with templates of 20-80% GC content Detects as little as 1 pg of lambda DNA or plasmid DNA and 100 pg of plant or animal genomic DNA Amplifies whole blood directly without nucleic acid extraction (EDTA, citrate, or heparin anticoagulants; dried blood spot cards) Tolerates bacterial culture up to 12.5% of the reaction volume and crude cDNA up to 15% of the reaction volume (50 uL total) Stable through 30 repeated freeze-thaw cycles |
| Specificity | High amplification specificity demonstrated across templates of 20-80% GC content, long human genomic fragments (1-40 kb), and direct amplification from bacterial cultures and whole blood. |
| Reaction Conditions / Protocol | Use a 2x master mix format; add only primers and template. Recommended annealing temperature: 60 degC. Extension speed: 10 sec/kb; for fragments above 10 kb, use 68 degC with 10 sec/kb; fragments below 10 kb can be amplified at 5 sec/kb. Whole blood preserved with EDTA, citrate, or heparin, and dried blood spot cards, can be amplified directly without extraction. |
| Components / Formulation | 2x master mix containing high-fidelity DNA polymerase, dNTPs, optimized long-range reaction buffer, and a proprietary protective agent (dye-free). |
| Storage Conditions | Store at -25 to -15 degC. Thaw slowly on ice; do not thaw at room temperature or by heating. |
| Shelf Life | 1 year when stored at -25 to -15 degC. |
| Package Specifications | 250 uL 1 mL 5x 1 mL 25x 1 mL |
| Product Form | Mix (2x ready-to-use master mix, no dye) |
| Quality Control | All reagent components undergo strict quality control to maximize product performance and batch-to-batch consistency; functional testing includes long-range amplification (up to 40 kb), fidelity measurement by next-generation sequencing, and sensitivity testing down to 1 pg lambda DNA. |
| Concentration | 2x |
| Source / Origin | Recombinant high-fidelity DNA polymerase formulation. |
| Expiration Date / Stability | Stable through 30 repeated freeze-thaw cycles; shelf life 1 year at -25 to -15 degC. |
| Compatibility | Suitable for templates with 20-80% GC content; amplifies whole blood (EDTA, citrate, or heparin preserved) and dried blood spot cards without extraction; tolerates bacterial culture up to 12.5% of the reaction and crude cDNA (including uracil-containing cDNA) up to 15% of the reaction. |
| Recommended Buffer System | Optimized long-range PCR buffer system (pre-mixed in the 2x master mix). |
| Application Notes / Precautions | For fragments above 10 kb, use 68 degC with an extension speed of 10 sec/kb to maximize success; fragments below 10 kb can be amplified at 5 sec/kb. For long cDNA amplification (1-14 kb), use a validated reverse transcription kit to prepare the cDNA template; when RNA is limiting or very long fragments are desired, skip digestion during reverse transcription and increase RNA/cDNA input. This is a high-fidelity (low error rate) product; if a mutation is observed in a product, re-amplify in replicate reactions to obtain a mutation-free product. High-GC fragments (20-80% GC) can be amplified; raising the temperature to 68 degC greatly improves the success rate for GC-rich targets. For direct amplification from bacterial culture, keep the culture at or below 12.5% of the reaction volume. Uracil-containing cDNA can be used as template; keep crude cDNA input within 15% of the reaction volume. Minimum detectable input: 100 pg plant or animal genomic DNA, 1 pg plasmid DNA. If a previously working reaction yields no band for long fragments: thaw the mix slowly on ice (never at room temperature or by heating), use 68 degC with 10 sec/kb, increase template amount for longer targets, and increase primer concentration or cycle number if needed. |
| Batch-to-Batch Consistency | All reagent components undergo strict quality control to maximize batch-to-batch stability; a proprietary protective agent maintains stable activity after repeated freeze-thaw (up to 30 cycles). |
For research use only, not for clinical use.
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