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| Product Name | Blood Advanced Direct PCR Kit |
| Catalog No. | RESK-0057 |
| Description | The Blood Advanced Direct PCR Kit allows direct PCR amplification from whole blood samples without DNA purification or any sample pretreatment. It is compatible with fresh, refrigerated, and frozen blood collected with common anticoagulants such as EDTA, heparin, and citrate, as well as commercial dried blood spots on Whatman 903 and FTA cards. The kit contains a genetically engineered DNA polymerase blend that combines high fidelity with strong tolerance to PCR inhibitors. The 2x Blood Advanced PCR Buffer is fortified with a potent processivity-enhancing factor, amplification enhancers, and an optimized buffer system, enabling efficient amplification of genomic fragments up to 8 kb under fast extension conditions; for fragments below 2 kb, an extension time of only 3-5 sec/kb is sufficient, dramatically shortening the time required for blood sample identification and testing. Amplification products generated by the Advanced High-Fidelity DNA Polymerase Mix have blunt ends, making them suitable for one-step rapid cloning and TOPO cloning workflows. The included Positive Control Primer Mix (10 uM each) amplifies a 237 bp fragment from a conserved sequence upstream of the sox21 gene in mammals and most vertebrates, providing a convenient positive control. |
| Intended Use | For direct PCR amplification from whole blood samples without DNA extraction or purification. Suitable for blood direct PCR, 3-4 plex multiplex PCR, and genotyping/identification of blood-derived samples. |
| Principle / Technology | The kit uses a genetically engineered, inhibitor-tolerant high-fidelity DNA polymerase blend together with a 2x blood-specific PCR buffer containing a processivity-enhancing factor and amplification enhancers. This system resists PCR inhibitors present in whole blood and supports fast, hot-start amplification with blunt-end products. |
| Sample Type | Fresh, refrigerated, or frozen whole blood anticoagulated with EDTA, heparin, or citrate; commercial dried blood spots on Whatman 903 and FTA cards (approx. 1 mm2 punch). Blood from multiple species (human, mouse, goat, chicken, pig, etc.) is supported. |
| Performance Range / Specifications | Amplifies genomic fragments up to 8 kb directly from whole blood. Extension rate of 3-5 sec/kb for fragments below 2 kb. Tolerates blood template concentrations from 0.5% up to 50% of the reaction (10% recommended). Compatible with templates of 30-75% GC content. Supports 3-4 plex multiplex PCR. Annealing temperature 60-(1-2) C. Hot-start enzyme; blunt-end products; no electrophoresis tracking dye. |
| Specificity | The included Positive Control Primer Mix (10 uM each) amplifies a 237 bp fragment from a conserved sequence upstream of the sox21 gene in mammals and most vertebrates, demonstrating broad species applicability across different anticoagulant-treated whole blood and blood cards. |
| Reaction Conditions / Protocol | Add whole blood directly to the PCR reaction at up to 50% of the total volume (10% recommended); no lysis or purification step is required. Use hot-start cycling with an annealing temperature of 60-(1-2) C and an extension time of 3-5 sec/kb for fragments below 2 kb (10 sec/kb may be used for longer or high-GC targets). |
| Components / Formulation | 2x Blood Advanced PCR Buffer Advanced High-Fidelity DNA Polymerase Mix Positive Control Primer Mix (10 uM each) |
| Storage Conditions | Store at -20 C. |
| Shelf Life | 1 year when stored at -20 C. |
| Package Specifications | 20 T; 50 T; 100 T; 500 T |
| Product Form | Kit |
| Quality Control | Kit performance is verified by direct amplification of DNA fragments of different lengths (up to 8 kb) from human whole blood, and by amplification of the 237 bp positive control fragment from whole blood and blood cards of multiple species and anticoagulant types. |
| Concentration | 2x (PCR buffer supplied as a 2x concentrate; positive control primers at 10 uM each) |
| Shipping Conditions | Shipped on dry ice. |
| Expiration Date / Stability | Valid for 1 year when stored at -20 C. |
| Compatibility | Compatible with EDTA-, heparin-, and citrate-anticoagulated whole blood (fresh, refrigerated, or frozen) and with dried blood spots on Whatman 903 and FTA cards. Blunt-end products are compatible with one-step rapid cloning and TOPO cloning kits. |
| Application Notes / Precautions | No product or low yield: check primer design, concentration, and purity; raise the annealing temperature or run an annealing gradient; increase the cycle number; try different blood input volumes. High-molecular-weight smear: avoid excessive extension times; reduce cycle numbers; lower primer concentration; try different blood input volumes. Low-molecular-weight smear: optimize the annealing temperature; reduce cycle numbers; lower primer concentration; redesign primers. Centrifuge the reaction and load the supernatant before electrophoresis; repeat the experiment to confirm the reaction setup and program. |
For research use only, not for clinical use.
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