AffiCLONE® II One Step Cloning Kit
The AffiCLONE® technology is a simple, fast, and highly efficient DNA seamless cloning technology. It enables directional cloning of inserts into any site in any vector. Use any method to linearize the vector, and introduce the end sequence of the linearized vector at the 5' end of the insert forward/reverse amplification primer, so that the 5' and 3' ends of the PCR product have the same ends sequence (15 - 20 bp) as the linearized vector, respectively. The PCR product and the linearized vector are mixed in a certain proportion. Under the catalysis of recombinase, the transformation can be performed at 37°C for 30 min to complete the directional cloning. ClonExpress II is a new generation of recombinant cloning kit. It has a unique ligase-independent system, which significantly reduces the self-ligation background of the vector, and the positive rate can reach more than 95%. Highly optimized reaction buffer and enhanced recombinase Exnase II can significantly improve the recombination efficiency and the tolerance to impurities. These features make it possible to use linearized vectors and inserts directly for recombinant cloning without purification, which significantly simplifies the experimental steps.
Components:
| Components | 25 Reactions | 50 Reactions |
| 5X CE II Buffer | 100 μL | 200 μL |
| Exnase II | 50 μL | 100 μL |
| 500 bp control insert (20 ng/μl) | 5 μL | 5 μL |
| pUC19 control vector，linearized (50 ng/μl，Amp+) | 5 μL | 5 μL |