| abstract |
The invention belongs to the field of biotechnology, and in particular relates to a one-step method for assembling BbsI enzyme cleavage and connecting fragments, an assembling kit and applications. After extensive and in-depth research, through a lot of screening and exploration, this application proposes for the first time a one-step BbsI digestion and ligation assembly kit and assembly method, which combines BbsI restriction endonuclease digestion with T4 DNA polymerase For one, the steps of cloning construction are greatly reduced, which can be applied to the assembly of short fragments such as coding sgRNA, and greatly reduce the operation process and steps. Compared with other assembly or construction kits, the present invention has high assembly efficiency, can be used for connection and assembly of different lengths and multiple fragments, and is suitable for assembly and construction of multiple modules. |