Product Overview
FireGene One Step Cloning technology is a Fast, Efficient, and simple DNA directional cloning technology, which can directionally clone the PCR product of the target gene to any site of any vector. FireGene One Step Cloning Kit is a new generation recombination cloning kit with the core of recombinase obtained by heterologous expression of a gene of T4 phage. UvsXase is compatible with conventional digestion and PCR reaction systems. Cloning vector restriction products or PCR products and target gene PCR products can be directly used for recombinant cloning without DNA purification, which greatly simplifies the experimental steps.
Applications
- DNA site-directed mutagenesis
- High-throughput cloning
- Seamless cloning
- Fast cloning
Components
Component Name | Amount | Storage (°C) |
UvsXase | 50 μl | -20 |
5 xUFO Buffer | 100 μl | -20 |
Experiment Workflow
Basic Steps:
- Amplify the whole plasmid of the vector at the cloning site to obtain a linearized vector;
- Design homologous primers so that the 5' and 3' ends of the PCR product of the target fragment have the completely identical sequence as the front and rear ends of the pre-inserted linearized vector, respectively (15 bp ~ 20 bp);
- After mixing the amplified PCR product of the target gene and the linearized vector in a certain proportion, under the catalysis of UvsXase, the transformation can be carried out within 30 minutes, and the directional cloning is completed.
The positive rate of clones can reach more than 95%.
Restrictions
For Research or Educational Use Only.