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| Key Features: | • Simple: one-tube super mix. • Fast: cloning within 5 - 15 min. • Efficient cloning of fragments of 50 bp-10 kb, with positive clone rate > 95%. • Directional cloning at any site on any vector, no limits in restriction enzyme choice. • Suitable for cloning of both single and multiple DNA fragments. |
| Description: | LiClone™ One Step Cloning Kit is simple, fast, and high efficient reagent which enables directional insertion of any amplified DNA product into any linearized vector at any site. Firstly, the vector is linearized at the cloning site. A small sequence overlapped with each end of the cloning site is added onto the insert through PCR. The insert and the linearized vector, with overlapped sequences of 15 bp - 20 bp on both 5’- and 3’-end, respectively, are mixed in an appropriate ratio and incubated with recombinant Exnase at 50℃ for 5 - 15 min. The Kit is applicable for homologous recombination with 1 - 4 fragments. The kit is independent of DNA ligase, significantly reducing the self-ligated colonies. The highly optimized 5× LiClone Master Mix significantly improve the recombination efficiency and the tolerance to impurities, enabling the insert to be used directly for recombination without any treatments to simplify the procedure. |
| Applications: | • Fast Cloning • High-throughput Cloning • Seamless Assembly • Site-specific Mutagenesis |
| Component: | 1. 5× LiClone Master Mix: 50 µl |
| Storage: | All components should be stored at -20°C; Avoid repeated freezing and thawing. |
| Documents: | Manual |
Case Study
LiClone™ One Step DNA Assembly Kit can clone 50bp-10kb fragment, single fragment with different length (50bp, 2kb, 4.5kb, 7.5kb, 10kb), multiple fragments: four fragments (3kb), three fragments (10kb), five fragments (3kb) and five fragments (6kb) with high efficiency; after purification can be homologously recombinated with pCE-Zero vector; recombination products will be used to transform competent cell DH5α. This kit is independent with ligase, self-link without vector and the positive cloning rate can be as high as 95% or above. After transformation, 50 clones are identified by PCR, the positive rates of insertion with different fragment length are shown right.
