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| Product Name | 2× PCR Master Mix for PAGE |
| Catalog No. | RESK-0053 |
| Description | This 2× PCR Master Mix for PAGE contains a ligand-modified, hot-start thermostable Taq DNA polymerase fused with a 3'-5' proofreading factor, together with an optimized buffer system that provides outstanding inhibitor tolerance and amplification efficiency. The reagent already includes the dNTPs and Mg2+ required for amplification; only primers and template need to be added, eliminating pipetting losses during setup. A tracking dye is premixed into the mix, so PCR products can be run directly on both agarose and polyacrylamide gels. Stabilizing agents in the formulation keep the product active through repeated freeze-thaw cycles. For added convenience, the mix can be paired with high-resolution precast PAGE gels. |
| Intended Use | PCR amplification specifically intended for downstream polyacrylamide gel electrophoresis (PAGE) analysis; also fully compatible with agarose gel electrophoresis. |
| Principle / Technology | A ligand-modified hot-start thermostable Taq DNA polymerase fused with a 3'-5' proofreading factor is combined with an optimized buffer containing dNTPs, Mg2+ and a tracking dye. The buffer is specifically formulated to avoid the band distortion and high silver-staining background that conventional PCR buffers cause on PAGE gels. |
| Sample Type | cDNA (e.g., human 293 cell cDNA), human genomic DNA, lambda DNA. |
| Performance Range / Specifications | Supports both PAGE and agarose gel electrophoresis Amplicon length: up to 5 kb Extension time: 10 sec/kb Recommended annealing temperature: 60°C High specificity, fast amplification and high detection rate from 100 ng template |
| Sensitivity / LOD | Detects as little as 50 fg of template (832 bp fragment amplified from dilution series of lambda DNA). |
| Specificity | High amplification specificity demonstrated on both cDNA and genomic DNA templates, verified by agarose and PAGE analysis. |
| Reaction Conditions / Protocol | Add primers and template directly to the 2× master mix. Recommended annealing temperature: 60°C. Extension time: 10 sec/kb (e.g., 8 s for an 832 bp fragment). Load the finished reaction directly onto agarose or polyacrylamide gels. |
| Components / Formulation | Ligand-modified hot-start thermostable Taq DNA polymerase fused with a 3'-5' proofreading factor dNTPs Mg2+ Optimized PAGE-compatible reaction buffer Tracking dye Stabilizing (protective) agents |
| Storage Conditions | Store at -20°C. |
| Shelf Life | 1 year when stored at -20°C. |
| Package Specifications | 1 mL 5× 1 mL 50× 1 mL |
| Product Form | Mix |
| Concentration | 2× |
| Shipping Conditions | Shipped with ice packs. |
| Expiration Date / Stability | Stable for 1 year at -20°C; the formulation remains active after repeated freeze-thaw cycles. |
| Compatibility | Optimized for polyacrylamide gel electrophoresis (including silver-stained PAGE) and fully compatible with agarose gel electrophoresis; can be paired with high-resolution precast PAGE gels. Products carry 3'-dA overhangs. |
| Application Notes / Precautions | Conventional PCR mixes are unsuitable for PAGE: their buffer components distort bands during polyacrylamide gel electrophoresis, and silver ions used in PAGE silver staining react with the buffer to produce strong background. This mix is specifically formulated to avoid both problems. |
For research use only, not for clinical use.
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