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.
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).
Figure 1. Experimental workflow of single fragment insertion into a vector using the GeneCopoeia Fast-Fusion™ Cloning Kit
Figure 2. PCR screening results. Lane1-3: 500 bp, lane 4-6: 1 kb, lane 7-9: 2 kb, lane 10-12: 4 kb, lane 13-15: 6 kb, lane 16-18: 8 kb, lane 19-21: 11 kb. Seven DNA fragments of different lengths were cloned into same vector respectively by Fast-Fusion Cloning Kit. Colonies were selected randomly for PCR screening after transformation. The results showed that all reactions gave high efficiency.