In-Fusion Cloning technology for high-throughput, seamless, and rapid cloning

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.

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.

The In‑Fusion Snap Assembly Master Mix 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 Snap Assembly Master Mix comes in either 500-μl liquid aliquots (standard format) or prealiquoted, lyophilized in 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.

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Overview

In‑Fusion Cloning enables directional, seamless cloning of any PCR insert(s) into any linearized vector in a single-tube, one-step, 15-minute reaction. No additional treatment of the PCR product (e.g., restriction digestion, ligation, phosphorylation, or blunt-end polishing) is required before proceeding to the cloning reaction.

Subcloning is unnecessary—Clone any insert, at any locus, directly into any final destination vector
Highly efficient—Over 95% efficiency has been demonstrated with a broad range of fragment sizes, from 0.5 to 15 kb
Seamless construction—Final constructs have no extra base pairs left over (as is often the case with restriction digests or TA cloning)
Flexibility to clone single or multiple fragments—Multiple inserts can be simultaneously cloned just as easily as single inserts, in just a single cloning reaction
Can be used to perform site-directed mutagenesis—No need to buy a separate kit; click here to learn more

Reagent configurations for high-throughput In-Fusion Cloning kits
Kit	Size	In‑Fusion Snap Assembly Master Mix	Stellar Competent Cells
In‑Fusion Snap Assembly Master Mix	250 rxns	500 μl	N/A
In‑Fusion Snap Assembly Master Mix	500 rxns	2 x 500 μl	N/A
In‑Fusion Snap Assembly Master Mix	1,000 rxns	4 x 500 μl	N/A
In‑Fusion Snap Assembly EcoDry	96 rxns	Prealiquoted, 96‑well plate	N/A
In-Fusion Snap Assembly EcoDry with Competent Cells	96 rxns	Prealiquoted, 96‑well plate	96‑well plate (20‑μl aliquots)

For custom bulk and dispense inquiries, contact your local Territory Manager or submit an online inquiry.

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Applications

High-throughput (HTP) cloning
PCR cloning
Multiple-fragment cloning
Gene synthesis
Gene design
Mutagenesis
Domain swapping
Domain modification

Additional product information

Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.

In-Fusion Cloning for high-throughput antibody development

A rapid, high-throughput cloning workflow for antibody development

Generating monoclonal antibodies requires efficient and rapid protocols for cloning, expression, and screening. Traditional cloning creates bottlenecks in this research, as it is often hampered by lengthy protocols, low efficiency, and reading frame errors. See how one lab has streamlined the process with In-Fusion technology, a highly accurate, seamless cloning method which is easily adaptable to high-throughput workflows.

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Streamline your plasmid purification workflow

NucleoSpin 96 Plasmid & NucleoSpin 96 Plasmid Transfection-grade

Time-saving parallel isolation of plasmid DNA from up to 5 ml of bacterial culture per prep
Protocol can be fully automated on liquid-handling platforms using vacuum processing
Innovative MN Wash Plate minimizes risk of cross contamination
Transfection-grade kit includes novel buffer technology for efficient endotoxin removal

Standard Transfection grade

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