Introduction
For rapid and effective cloning of PCR products
The GeneCopoeia Fast-Fusion™ Multi Seamless Cloning Kit provides a rapid method for cloning your PCR products. In as little as 5 minutes minutes at 50℃, any PCR fragment can be cloned into your linearized vector. After a simple clean up step, a PCR-generated DNA fragment or other purified DNA fragment can be joined to a vector with overlapping ends (Fig.A).Up to five DNA fragments can be joined together in a single reaction. A single fragment can be efficiently fused and assembled in 5 minutes. Well-prepared vectors generate almost 100% positive clones.
No restriction sites are needed at the junction, enabling insertion of your fragment of interest at any vector position. The linearized vector can be generated by either PCR or restriction enzyme digestion. The PCR products can be produced by either Taq DNA polymerase or other high fidelity DNA polymerase.
Advantages
- Fast and simple—1 minute for operation and5 minutes for incubation.
- High adaptability—No restriction or recombination sites needed, insert fragments generated by either PCR or restriction enzyme digestion can be used.
- Seamless construction—Final constructs are seamless with no extra or unwanted base pairs.
- Flexibility—Multiple inserts can be assembled in one reaction. Suitable for multi-site mutagenesis.
- High efficiency—Greater than 90% of colonies after transformation contain the correct insert(s).
Portocol overview
Figure A. Experimental workflow of using the Fast-FusionTM Multi Seamless Cloning Kit
