ClonExpress® technology is a fast, simple and efficient DNA seamless cloning technology, enabling the insert of any position of any vector to be cloned. The user can linearize the vector and place the insert 5' in the forward/reverse PCR primer.
Then, the end sequence of the linearized vector is introduced into the end, so that the 5' and 3' ends of the PCR product have the same sequence as the two ends of the linearized vector (15 bp-20 bp).
This enables the linearized vector and the PCR product to be in a specific ratio. Post mixing, under the catalysis of the recombinase, the reaction is carried out for 30 minutes at 37 °C to complete the directional cloning.
ClonExpress® II, a novel generation recombinant cloning kit, possesses a unique non-ligase-dependent system, thus drastically reducing the background of vector self-ligation and enabling the positive rate to reach over 95%.
The highly optimized reaction buffer and the improved recombinase Exnase II enhances the tolerance to impurities and the recombination efficiency of cloning, thus enabling the direct use of linearized inserts and vectors for recombination cloning. This avoids purification and simplifies the experimental steps.
Storage conditions
Product advantages
- Avoids the need to consider the restriction site carried by the insert
- Quick, simple, efficient and suitable for any carrier
- Efficient cloning of 50 bp–10 kb DNA fragments
- No self-connection, no ligase, positive rate >95%
- PCR products and linearized cloning vector can be cloned directly without purification
Applications
- High-efficiency cloning
- Quick cloning
- Site-specific mutagenesis
- Seamless assembly