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| Product Name | 2× AdvanceFast High-Fidelity PCR Master Mix (With Dye) |
| Catalog No. | RESK-0031 |
| Description | The 2x AdvanceFast High-Fidelity PCR Master Mix (With Dye) is a ready-to-use 2x premixed solution containing an AdvanceFast high-fidelity DNA polymerase, dNTPs, and an optimized buffer system. A loading dye is pre-mixed into the master mix, so PCR products can be loaded directly onto agarose gels for electrophoresis. Amplification products are blunt-ended. The master mix is fast and easy to use - only primers and template need to be added - and offers high sensitivity, strong specificity, and excellent stability. A proprietary protective agent keeps the premix active even after repeated freeze-thaw cycles. |
| Intended Use | Fast, high-fidelity PCR amplification of genomic DNA, cDNA, and crude samples (blood, tissue lysates, bacterial cultures); products are ready for direct gel electrophoresis; for research use. |
| Principle / Technology | An AdvanceFast high-fidelity DNA polymerase with proofreading activity delivers very low error rates (231 mismatches per 300,000 bases in GC-rich, mutation-prone sequences) at extension speeds as fast as 5 sec/kb. The premixed loading dye enables direct gel electrophoresis. Products are blunt-ended with no A overhang. |
| Sample Type | Genomic DNA (human, mouse, plant species such as Arabidopsis, rapeseed, potato, maize, and rice; animal models such as Drosophila, C. elegans, and zebrafish), cDNA, bacterial culture, whole blood (direct amplification, up to 20% of the reaction), and mouse lysate. |
| Performance Range / Specifications | High fidelity: mismatch/mutation rate of 231 per 300,000 bases in GC-rich, mutation-prone sequences (fidelity 83x) Fast extension: as fast as 5 sec/kb for fragments up to 10 kb Amplification length: human gDNA/cDNA up to 10 kb; lambda DNA up to 16 kb; rice gDNA fragments up to 20 kb demonstrated Covers templates with 20-80% GC content (validated for 23-76% GC targets from human gDNA) Tolerates 1-20% blood in the reaction and shows resistance to inhibition by mouse lysate Stable for 7 days at 37 degC without loss of performance Products are blunt-ended, compatible with downstream homologous recombination cloning and TOPO blunt-end cloning |
| Specificity | Excellent amplification specificity demonstrated across 0.4-20 kb fragments from diverse templates (human, mouse, plant, and bacterial) with 20-80% GC content, and high detection rates for randomly selected sequences from multiple species. |
| Reaction Conditions / Protocol | Use a 2x master mix format; add only primers and template. Recommended annealing temperature: 60 degC. Extension speed: 5 sec/kb (5-10 sec/kb depending on template and fragment length). Products carry a pre-mixed loading dye and can be loaded directly onto agarose gels for electrophoresis. |
| Components / Formulation | 2x master mix containing AdvanceFast high-fidelity DNA polymerase, dNTPs, optimized reaction buffer, loading dye, and a proprietary protective agent. |
| Storage Conditions | Store at -25 to -15 degC. |
| Shelf Life | 1 year when stored at -25 to -15 degC. |
| Package Specifications | 1 mL 5x 1 mL |
| Product Form | Mix (2x ready-to-use master mix with loading dye) |
| Quality Control | Fidelity validated by cloning and sequencing of 100 single colonies per target across 6 GC-rich, mutation-prone fragments; performance confirmed across multiple species, GC contents (20-80%), and template types. |
| Concentration | 2x |
| Source / Origin | Recombinant high-fidelity DNA polymerase formulation. |
| Expiration Date / Stability | Stable for 7 days at 37 degC without affecting performance; a proprietary protective agent maintains activity after repeated freeze-thaw; shelf life 1 year at -25 to -15 degC. |
| Compatibility | Tolerates 1-20% blood and mouse lysate; compatible with templates of 20-80% GC content from human, animal, plant, and bacterial sources. Blunt-end products are suitable for homologous recombination cloning and TOPO blunt-end cloning; for TA cloning, dATP and a conventional Taq polymerase are required separately. |
| Recommended Buffer System | Optimized fast high-fidelity PCR buffer system (pre-mixed in the 2x master mix). |
| Application Notes / Precautions | The With Dye version contains a pre-mixed loading dye, eliminating the need to purchase and add loading buffer well by well before gel electrophoresis; a dye-free version is recommended if downstream restriction digestion is planned, to avoid interference and ensure efficient digestion. Amplification products carry no A overhang (blunt ends) and can be used directly for homologous recombination cloning or TOPO blunt-end cloning; for TA cloning, add dATP tailing with a conventional Taq polymerase (purchased separately). |
| Batch-to-Batch Consistency | A proprietary protective agent maintains stable activity after repeated freeze-thaw; the formulation remains fully functional after 7 days at 37 degC, supporting reproducible performance. |
For research use only, not for clinical use.
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