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 and incubated with Exnase II at 37℃ for 30 min (Fig. 1.). The cloning products can then be directly transformed to competent cells with a positive rate > 95%.
ClonExpress MultiS One Step Cloning Kit is a new version cloning kit based on ClonExpress One Step Cloning technology. Exnase MultiS and reaction buffer in this kit are especially optimized for the assembly of multiple fragments. With the help of this kit, up to five fragments can be orderly assembled in a single reaction. In addition, Exnase MultiS is compatible with the reaction systems of restriction enzyme digestion and PCR, which means that both the digestion products of vector and the PCR products of insert can be directly used for recombination without purification, significantly simplifying the procedures.
Key Advantages:
• Easy, fast, and efficient.
• Directional cloning at any site on any vector.
• No need to consider the restriction enzyme cutting sites carried on inserts.
• Orderly assembly of up to five fragments simultaneously.
• Linearized vector and PCR products can be used directly without purification.
Cloning competent cells can be ordered with this cloning kit. Please ask us for help if you need to add the competent cells to your order of this cloning kit.