Seamless cloning technique from HANBIO is a new, fast and simple cloning method, which can insert one or more target DNA fragments at any site of the plasmid without any restriction enzymes and ligases. Breaking through the traditional double enzyme digestion and linking, the recombinant vector of high efficiency cloning can be obtained by one-step recombinant method, which greatly improves the working efficiency.

Introduction of Seamless Cloning Kit

The HB-infusion™ seamless cloning kit developed by HANBIO is extremely simple to operate. It only needs to linearize the cloning site of the vector and introduce 15-25 bp homologous sequences that are completely consistent with both ends of the vector cloning site at the 5 'end of the insert PCR primer. The linearized cloning vector and the PCR fragments with homologous sequences were mixed in appropriate proportions and then added into HB-infusion™ Master mix. The directed cloning could be completed quickly by the reaction of DNA exonuclease, DNA polymerase and ligase in the reaction system at 50℃ for 20 minutes. The positive rate was almost 100%.

Features of Seamless Cloning Kit

The Operation is Simple and Fast: The seamless cloning kit from HANBIO does not require any restriction enzymes, ligases, phosphorylation, terminal complementation, etc. The reaction time of the system was only 20 minutes.

Wide Application: Multiple DNA fragments, up to 10, can be linked simultaneously.

Precise and Efficient: No extra sequence is attached, precise directional connection, high positive rate.

Infusion Cloning Principle

The design of primers for gene cloning and the amplification of target DNA fragments are the same as conventional PCR. The only difference is that the end of the vector and the end of the primer should have 15 to 20 homologous bases, and the resulting PCR product will carry 15 to 20 homologous bases with the sequence of the vector.

Seamless Cloning VS Traditional Cloning

Seamless Cloning Kit Product Information

Code No. Component Usage Counter Volume

HB-infusion-20T 2 x HB-infusion™ Master mix 20 tests 200µL

Positive linearized pUC vector （250 ng） 5 tests 25µL

Positive control insert (500 ng) 5 tests 25µL

Storage condition -20℃