In-Fusion HD EcoDry Cloning Plus kits are provided in a convenient lyophilized, ready-to-use format that can be stored at room temperature. All the components for the cloning reaction are pre-aliquoted and dried down in tubes that are covered with an aluminum seal. Minimal handling eliminates chances of contamination, pipetting errors, or missing reagents. In-Fusion EcoDry Cloning Plus Systems allow you to clone any PCR fragment into any linearized vector in a single step without restriction digestion of the PCR fragment, ligation, or blunt-end polishing. The ability to clone directly into any vector at any restriction site eliminates the need for any further subcloning or manipulation.
How it Works
The In-Fusion Enzyme premix fuses PCR-generated sequences and linearized vectors efficiently and precisely, utilizing a 15 bp overlap at their ends. This 15 bp overlap can be engineered by designing custom primers for amplification of the desired sequences. This method can be used to clone single or multiple fragments into a single vector without subcloning. All that’s required is a simple 30-minute incubation.
In-Fusion HD Plus EcoDry Cloning Kit Components
All In-Fusion HD Plus EcoDry Cloning kits contain the In-Fusion HD enzyme premix and reagents for control experiments, in a lyophilized format that provides ease of use and consistency. The In-Fusion premix comes in ready-to-use microtubes that can be stored right on the lab bench until you're ready to perform the In-Fusion reaction. In-Fusion HD Plus EcoDry Cloning kits also include:
- CloneAmp HiFi High Fidelity PCR Premix, a convenient 2X master mix that provides exceptionally accurate, efficient and fast DNA amplification. The premix contains dNTPs and an optimized buffer, allows rapid set-up of PCR reactions, and facilitates successful cloning.
- Stellar Competent Cells, an HST08 E. coli strain that provides high transformation efficiency (greater than 5 x 108 cfu/µg) and compliments the efficiency of the In-Fusion HD Cloning Plus kit.
- NucleoSpin Gel and PCR Clean-Up, for PCR purification and gel extraction.