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| Product Name | Universal One-Step Cloning Kit |
| Catalog No. | RESK-0038 |
| Description | The Universal One-Step Cloning Kit is a next-generation homologous recombination cloning system designed for fast, seamless, and highly efficient DNA assembly. Its optimized 2× enzyme premix combines a recombinase, a ready-to-use reaction buffer, and a proprietary recombination enhancer that markedly boosts cloning efficiency. With this kit, PCR products can be directionally cloned into any site of any vector, even at vector concentrations as low as 5 ng/μL, and up to six fragments can be joined in a defined order in a single reaction. Simply linearize the vector, add 15-25 bp homology arms matching the linearized vector ends to the 5' ends of the insert PCR primers, and incubate the PCR products with the linearized vector at 50°C for 5-15 minutes. More than 95% of the resulting colonies are positive clones. |
| Intended Use | Seamless, directional cloning of 1-6 PCR fragments into any site of any vector via homologous recombination; ideal for single-insert cloning and ordered multi-fragment assembly in a single tube. |
| Principle / Technology | Homologous recombination. Inserts are amplified with primers carrying 15-25 bp 5' extensions homologous to the linearized vector ends, so the PCR products share identical terminal sequences with the vector. The recombinase in the 2× premix, assisted by a unique recombination enhancer, directionally joins the insert(s) and vector at 50°C without restriction digestion or ligation. |
| Sample Type | PCR products (purified or unpurified) and linearized cloning vectors |
| Performance Range / Specifications | Directional insertion of 1-6 DNA fragments Fragment length range: 50 bp-23 kb Positive clone rate: >95% for single-fragment cloning Multi-fragment ligation completed in a single tube Vector input as low as 5 ng per 20 μL reaction Ligation reaction completed in as little as 5 minutes (15-30 min recommended for multi-fragment assemblies) |
| Specificity | Directional cloning is driven by 15-25 bp terminal homology arms identical to the linearized vector ends, ensuring site-specific, orientation-specific insertion at any chosen vector locus. |
| Reaction Conditions / Protocol | 50°C for 5-15 minutes (30 minutes recommended for six-fragment assembly). Recommended vector:insert molar ratio 1:3; approximately 5 ng of linearized vector per 20 μL reaction. Inserts require 15-25 bp homology arms at the 5' ends of both PCR primers matching the linearized vector ends. |
| Components / Formulation | 2× Universal One-Step Cloning Enzyme Premix (recombinase, optimized reaction buffer, and recombination enhancer) |
| Storage Conditions | Store at -20°C. |
| Shelf Life | 1 year when stored at -20°C. |
| Package Specifications | 20 T, 50 T |
| Concentration | Compatible with vector concentrations as low as 5 ng/μL |
| Shipping Conditions | Shipped with ice packs. |
| Expiration Date / Stability | Stable for 1 year when stored at -20°C. |
| Compatibility | Compatible with any vector and any cloning site; accepts purified or unpurified PCR fragments; supports single-fragment and 2-6 fragment ordered assembly. |
| Recommended Buffer System | Optimized homologous recombination buffer, pre-mixed in the 2× enzyme premix. |
For research use only, not for clinical use.
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