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| Product Name | 2× Ultra Long-Range PCR Master Mix (With Dye) |
| Catalog No. | RESK-0061 |
| Description | This 2x master mix is built for long-range and high-accuracy PCR. It contains a ligand-modified thermostable Taq DNA polymerase fused with a 3'-5' proofreading factor in an optimized buffer system, enabling amplification of genomic fragments up to 25 kb, cDNA sequences up to 14 kb, and lambda DNA fragments up to 40 kb. The premix already contains the dNTPs and Mg2+ required for amplification, so only primers and template need to be added, avoiding pipetting losses during reaction setup. An electrophoresis loading dye is also included, greatly streamlining downstream gel analysis, and a protective agent keeps the mix active even after repeated freeze-thaw cycles. PCR products carry 3'-dA overhangs and can be ligated directly into T vectors. |
| Intended Use | For long-range (long-fragment) PCR amplification of genomic DNA, cDNA, and lambda DNA templates. |
| Principle / Technology | Ligand-modified hot-start thermostable Taq DNA polymerase fused with a 3'-5' proofreading (correction) factor, in an optimized buffer system; a protective agent maintains stable activity after repeated freeze-thaw cycles. |
| Detection Method | Endpoint PCR; products are analyzed by agarose gel electrophoresis (loading dye is included in the mix). |
| Sample Type | Genomic DNA from various species (plant and animal), cDNA, and lambda DNA. |
| Performance Range / Specifications | Amplification length: up to 25 kb genomic DNA, up to 14 kb cDNA, up to 40 kb lambda DNA Extension time: 30 s/kb Annealing temperature: 68°C Template GC content: 30-70% Product ends: 3'-dA overhangs, directly ligatable into T vectors Validated amplification of lambda DNA targets of 25, 30, 35, and 40 kb (50 ng template, 30 s/kb extension) Validated amplification of 10-20 kb genomic targets from corn, Arabidopsis, and mouse, and 14 kb human cDNA targets (RIF1 and MACF1 genes from 293 cell cDNA) |
| Sensitivity / LOD | A 14 kb DNA fragment was successfully amplified from as little as 1 ng of human genomic DNA (tested input range: 1-2000 ng). |
| Specificity | The ligand-modified hot-start polymerase and the fused proofreading factor provide high amplification specificity with a degree of fidelity. |
| Reaction Conditions / Protocol | 2x premix containing dNTPs and Mg2+; add only primers and template. Recommended extension time: 30 s/kb; recommended annealing temperature: 68°C. |
| Components / Formulation | 2x Ultra Long-Range PCR Master Mix (With Dye): ligand-modified hot-start Taq DNA polymerase, fused 3'-5' proofreading factor, dNTPs, Mg2+, electrophoresis loading dye, and protective agents in an optimized buffer |
| Storage Conditions | Store at -25 to -15°C. |
| Shelf Life | 1 year. |
| Package Specifications | 1 mL; 5 x 1 mL; 50 x 1 mL |
| Product Form | Mix (2x premix, with electrophoresis loading dye) |
| Concentration | 2x |
| Expiration Date / Stability | Valid for 1 year at -25 to -15°C; the protective formulation maintains stable activity after repeated freeze-thaw cycles. |
| Compatibility | Broad species compatibility (validated on corn, Arabidopsis, and mouse genomic DNA, and human 293 cell cDNA). PCR products with 3'-dA overhangs are directly compatible with T-vector (TA) cloning. |
For research use only, not for clinical use.
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