The classic method of using restriction enzyme digestion and ligation for vector construction is limited by the specific restriction sites on the vector. Therefore, when constructing multiple insertion fragments, multiple rounds of digestion and ligation are often required. This process is not only cumbersome but also introduces unwanted nucleotides, making it a relatively inefficient vector construction method.
Seamless cloning technology, based on the principle of recombination, relies on the recombination of homologous sequences of 15-25bp at the ends of the insertion fragments and the linearized vector. This method eliminates the relatively complex steps of digestion and ligation. Theoretically, it can clone insertion fragments to any site on any linearized vector, with high recombination efficiency and very low vector self-ligation background. It is a simple, fast, and efficient technology for directional DNA cloning.
Simple and Convenient: Compatible with PCR product systems and vector digestion systems, eliminating the need for tedious purification steps
High Positive Rate: Directional cloning of a single DNA fragment to the vector with a positive rate higher than 95%
Single or Multiple Fragments Connection: Simultaneous insertion of single or multiple fragments, eliminating repetitive purification and digestion processes, providing maximum flexibility
Simple Operation
Hanbio's HB-infusion™ seamless cloning kit is extremely simple to use. You only need to linearize the vector cloning site and introduce the 15-25bp homologous sequences to both ends of the vector cloning site at the 5' end of the insertion PCR primers.
Quick Completion
Mix the linearized cloning vector with the PCR fragment containing homologous sequences at an appropriate ratio, add them to Hanbio's seamless cloning kit, and through the action of exonuclease, DNA polymerase, and ligase in the reaction system, directional cloning can be quickly completed in 20 minutes at 50°C, with an almost 100% positive rate.
