Introduction
For rapid and effective cloning of PCR products
Fast-Fusion™ Cloning Kit provides a rapid method for cloning your PCR product. In just 15 minutes at room temperature, any PCR fragment can be cloned into your linearized vector at will. 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.1). Up to eight DNA fragments can be joined together in a single reaction. Well-prepared vectors generate almost 100% positive clones. There is no restriction site required at the junction site. Therefore, your fragment of interest can be inserted at any position in the vector. 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 and 15 minutes for incubation at room temperature.
- No restriction site or recombination site needed, insert fragments generated by either PCR or restriction enzyme digestion can be used.
- Final constructs have no extra base pairs remaining.
- Multiple inserts can be assembled in one reaction. Suitable for multi-site mutagenesis.
- Greater than 90% of colonies after transformation contain the correct insert(s).