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| Product Name | Veritas Long-Range High-Fidelity PCR Master Mix (With Dye) |
| Catalog No. | RESK-0030 |
| Description | The Veritas Long-Range High-Fidelity PCR Master Mix (With Dye) is a ready-to-use 2x premixed solution containing a high-fidelity DNA polymerase, dNTPs, and an optimized buffer system developed for long-range amplification. A blue electrophoresis tracking dye is pre-mixed into the master mix, so PCR products can be loaded directly onto agarose gels without adding loading buffer. Amplification products are blunt-ended. The master mix is fast and easy to use - only primers and template need to be added - and delivers high sensitivity, strong specificity, and excellent stability. A proprietary protective agent keeps the premix active even after repeated freeze-thaw cycles. |
| Intended Use | Long-range, high-fidelity PCR amplification of DNA templates (genomic DNA, cDNA, plasmids, and crude samples such as bacterial cultures), with direct gel loading of products; for research use. |
| Principle / Technology | A high-fidelity DNA polymerase with proofreading (3' to 5' exonuclease) activity combined with an optimized long-range buffer system enables accurate, rapid synthesis of long amplicons. The premixed blue tracking dye allows direct gel electrophoresis of PCR products. Products are blunt-ended. |
| Sample Type | Genomic DNA (human, animal, plant), cDNA (including uracil-containing cDNA with residual RNA), plasmid DNA, lambda DNA, and bacterial culture (direct amplification). |
| Performance Range / Specifications | Amplifies DNA fragments up to 40 kb and cDNA fragments of 1-14 kb Extension speed: 5-10 sec/kb (5 sec/kb for fragments below 10 kb; 10 sec/kb for fragments above 10 kb) Fidelity: 83x that of Taq DNA polymerase (measured by next-generation sequencing) 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 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. |
| Reaction Conditions / Protocol | Use a 2x master mix format; add only primers and template. Recommended annealing temperature: 60 degC. For fragments below 10 kb, use an extension speed of 5 sec/kb; for fragments above 10 kb, use 68 degC with 10 sec/kb. Products can be loaded directly onto agarose gels for electrophoresis. |
| Components / Formulation | 2x master mix containing high-fidelity DNA polymerase, dNTPs, optimized reaction buffer, blue electrophoresis tracking dye, and a proprietary protective agent. |
| 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 with blue dye) |
| Quality Control | All reagent components undergo strict quality control to ensure optimal 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; 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 | A proprietary protective agent maintains stable activity after repeated freeze-thaw (up to 30 cycles), ensuring reliable, reproducible performance between batches. |
For research use only, not for clinical use.
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