In-Fusion Cloning offers one-step, high-efficiency, and high-throughput cloning for projects that require large numbers of cloning reactions, such as expression screening studies and library construction.
Advancements in recombinant antibody-based therapeutic development highlight a need for streamlined, high-throughput workflows that allow rapid generation and screening of functional clones.
Depending on your own high-throughput needs, you may want to choose kits based on reagent configuration.
In-Fusion Cloning offers one-step, high-efficiency, and high-throughput cloning for projects that require large numbers of cloning reactions, such as expression screening studies and library construction.
Advancements in recombinant antibody-based therapeutic development highlight a need for streamlined, high-throughput workflows that allow rapid generation and screening of functional clones.
In‑Fusion Cloning offers a suitable method for rapid cloning of many constructs, with a workflow that is amenable to automation. In‑Fusion Cloning provides >95% efficiency, increasing the likelihood that the correct clones are obtained after one round of cloning.
Benefits of In‑Fusion Cloning over other methods for high-throughput applications
Unlike the Gibson cloning method, In‑Fusion Cloning does not rely on a low-fidelity polymerase to fill junction regions. This ensures that cloning done with the In-Fusion enzyme mix results in accurate sequences.
Conventional cloning methods (e.g., ligation-based cloning and TOPO cloning) introduce limitations such as extra subcloning steps, lower efficiency, and lack of insert directionality, making these methods incompatible with the needs of high-throughput cloning. In-Fusion Cloning is highly efficient, always directional, and suitable for use on automation platforms, thus eliminating bottlenecks associated with traditional cloning methods.
In-Fusion Cloning kits that include Cloning Enhancer (CE) provide an additional option for streamlining high-throughput workflows. CE is an easy-to-use enzyme premix that removes background plasmid DNA and PCR residue, eliminating the need for PCR purification prior to cloning. In-Fusion HD Cloning Plus CE kits are ideal for cloning when there is a single PCR product on the gel. CE is also available separately in 50-μl aliquots.
The In‑Fusion HD 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 sequence using our primer design tool.
The In‑Fusion HD cloning enzyme comes in either 100-μl liquid aliquots (standard format) or prealiquoted, lyophilized 96-well plates (EcoDry format) which are stable at room temperature, to accommodate robotic liquid handling. See the table below for specific reagent configurations for various kits.
