In-Fusion Snap Assembly Master Mix
In-Fusion Snap Assembly Master Mix
Brand: Takara Bio.
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In-Fusion Snap Assembly Master Mix
In-Fusion Snap Assembly master mixes for seamless DNA cloning
In-Fusion Snap Assembly products enable directional, seamless cloning of any PCR fragment—or multiple fragments—into any linearized vector in a single 15-minute reaction. No additional treatment of the PCR fragment (such as restriction digestion, ligation, phosphorylation, or blunt-end polishing) is required. The efficiency of In-Fusion Snap Assembly is over 95%, providing confidence in your clones and enabling you to scale up for high-throughput workflows.
The In-Fusion Snap Assembly Master Mix fuses PCR-generated sequences and linearized vectors efficiently and precisely—without the use of ligase—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 using our primer design tool. The In-Fusion master mix removes nucleotides from the 3' end of the linear DNA strands. This allows complementary base pairs between two pieces of DNA to anneal, leading to fragment joining. This method can be used to clone single or multiple fragments into a single vector without subcloning. Additional applications include mutagenesis, gene synthesis, gene design, domain swapping, and domain modification. The ability to clone directly into any destination vector at any locus (without the use of restriction enzymes) eliminates the need for any further subcloning or manipulation, simplifying the cloning workflow and further enabling a transition to high-throughput applications.
The In-Fusion Snap Assembly Master Mix is provided in a liquid format as a 5X enzyme premix in 20-μl to 500-μl aliquots, depending on kit size. It also contains reagents for control experiments.
Note: the 500-reaction and 1,000-reaction In-Fusion Snap Assembly Master Mixes contain two or four vials of the 250-reaction size, respectively.
Overview
In-Fusion Snap Assembly is a simple yet elegant cloning technology that has been optimized to give you exceptionally high accuracy, guaranteeing cloning success and saving you time and hassle.
- Subcloning is unnecessary: clone any insert directly into your final vector, regardless of your cloning locus
- Highly efficient: over 95% efficiency demonstrated with a broad range of fragment sizes, from 0.5 kb to 34 kb*
- Seamless construction: final constructs have no extra base pairs left over (as is often the case with restriction digest or TA cloning)
- Flexible experimental design: clone single or multiple DNA fragments simultaneously, with just a single reaction
- Site-directed mutagenesis: click here to learn how this streamlined technology can be applied to insertions, deletions, or substitutions
- *When used with Stellar Competent Cells, PrimeSTAR Max Polymerase, and NucleoSpin Gel and PCR Clean-up kit.
Applications
- PCR cloning
- High-throughput cloning (see full range of reagent options on our high-throughput cloning page)
- Multiple-fragment cloning
- Site-directed mutagenesis
- Gene synthesis
- Adding adaptors, linkers, and protein tags
- Cloning using modular parts
In-Fusion Cloning protocol for single-insert cloning.
In-Fusion Cloning protocol. Schematic of the standard workflow for all In-Fusion Cloning kits.
Performance comparison between In-Fusion Snap Assembly and NEBuilder HiFi using inverse PCR.
Performance comparison between In-Fusion Snap Assembly and NEBuilder HiFi using inverse PCR. A single 3.8-kb insert (Panel A) or a 34.2-kb adenovirus insert (Panel B) was cloned into a 2.7-kb vector which was linearized via inverse PCR. These cloning reactions were performed in triplicate with both In-Fusion Snap Assembly and NEBuilder HiFi. Primers were designed according to the manufacturers' specifications. After transformation and plating, 20 colonies from each replicate were analyzed by Sanger sequencing (for the 3.8-kb insert) or colony PCR (for the adenovirus insert) to determine the cloning accuracy. In-Fusion Snap Assembly yielded 2X more colonies than NEBuilder HiFi.
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In-Fusion Cloning protocol for single-insert cloning.
In-Fusion Cloning protocol. Schematic of the standard workflow for all In-Fusion Cloning kits.
Performance comparison between In-Fusion Snap Assembly and NEBuilder HiFi using inverse PCR.
Performance comparison between In-Fusion Snap Assembly and NEBuilder HiFi using inverse PCR. A single 3.8-kb insert (Panel A) or a 34.2-kb adenovirus insert (Panel B) was cloned into a 2.7-kb vector which was linearized via inverse PCR. These cloning reactions were performed in triplicate with both In-Fusion Snap Assembly and NEBuilder HiFi. Primers were designed according to the manufacturers' specifications. After transformation and plating, 20 colonies from each replicate were analyzed by Sanger sequencing (for the 3.8-kb insert) or colony PCR (for the adenovirus insert) to determine the cloning accuracy. In-Fusion Snap Assembly yielded 2X more colonies than NEBuilder HiFi.
