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| Product Name | 2× PCR Master Mix (With Dye) |
| Catalog No. | RESK-0049 |
| Description | The 2× PCR Master Mix (With Dye) is a ready-to-use premixed solution for routine PCR, containing Taq DNA polymerase, a dNTP mixture, MgCl2, and an optimized buffer system. Only primers and template need to be added, greatly simplifying reaction setup. The mix contains bromophenol blue dye, so PCR products can be loaded directly onto gels for electrophoresis. A high-quality stabilizer keeps the mix fully functional for up to 3 months at 4°C. PCR products carry 3'-dA overhangs and can be cloned directly into T vectors. |
| Intended Use | Routine (conventional) PCR amplification. |
| Principle / Technology | Standard endpoint PCR based on Taq DNA polymerase in an optimized buffer system. |
| Sample Type | Genomic DNA templates, including templates with high GC content (validated up to 70% GC, e.g., human 293T cell genomic DNA and Arabidopsis genomic DNA). |
| Performance Range / Specifications | Extension speed: 30-60 sec/kb. Amplicon length: up to 4-5 kb. GC compatibility: validated on templates with 40-70% GC content. Annealing temperature: Tm minus 2-5°C (60°C used in validation). Stability: full performance retained after 3 months at 4°C, 1 month at 25°C, or 1 year at -20°C. |
| Specificity | Reliably amplifies high-GC targets; validated on genes with 40%, 50%, 60%, and 70% GC content from human 293T cell genomic DNA at 60°C annealing and 30 sec/kb extension. |
| Reaction Conditions / Protocol | Annealing temperature: Tm minus 2-5°C (60°C recommended). Extension time: 30 sec/kb (30-60 sec/kb range). Add only primers and template to the 2× mix. |
| Components / Formulation | Taq DNA polymerase, dNTP mixture (200 µM each), MgCl2 (2 mM final concentration), optimized buffer system, bromophenol blue dye, and a high-quality stabilizer. |
| Storage Conditions | Store at -25°C to -15°C. |
| Shelf Life | 2 years when stored at -25°C to -15°C. |
| Package Specifications | 1 mL; 5× 1 mL; 50× 1 mL; 100× 1 mL |
| Product Form | Premixed solution |
| Quality Control | Performance verified by amplification of high-GC targets (40-70% GC) and by stability testing after 1 year at -20°C, 3 months at 4°C, and 1 month at 25°C. |
| Concentration | 2× |
| Source / Origin | Taq DNA polymerase. |
| Expiration Date / Stability | Stable for 2 years at -25°C to -15°C. Contains a high-quality stabilizer; full performance retained after 3 months at 4°C, 1 month at 25°C, or 1 year at -20°C. |
| Compatibility | Contains bromophenol blue dye for direct gel loading; PCR products carry 3'-dA overhangs suitable for TA cloning; validated on templates with up to 70% GC content. |
| Application Notes / Precautions | Not recommended for PAGE electrophoresis, as components in the mix may interfere with polyacrylamide gels. Reaction volumes can be scaled down proportionally, reducing primers and template accordingly. This With Dye version contains bromophenol blue for direct gel loading; the No Dye version requires adding DNA loading buffer before electrophoresis. Both dye-containing and dye-free products are suitable for downstream sequencing, since sequencing providers purify the PCR products first. The mix contains Mg2+ at a 2 mM final concentration and 200 µM of each dNTP. Direct restriction digestion of PCR products is not recommended, as some components may affect digestion efficiency. The enzyme has no 3'-5' exonuclease activity; products carry 3'-dA overhangs and are suitable for TA cloning. |
| Batch-to-Batch Consistency | Formulated with a high-quality stabilizer; consistent amplification performance maintained after 3 months at 4°C. |
For research use only, not for clinical use.
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