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.  
 

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.
 

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.