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The Seamless Cloning Master Mix is a ready-to-use reaction mix for seamless cloning reactions. It allows for successful assembly of multiple DNA fragments, regardless of fragment length or end compatibility. It has been rapidly adopted by the synthetic biology community due to its ease-of-use, flexibility and suitability for large DNA constructs.
The Seamless Cloning Master Mix efficiently joins multiple overlapping DNA fragments in a single-tube isothermal reaction. The system allows the cloning of the DNA fragments into virtually any linearized Escherichia. coli vector, requires no preexisting recombination sites or extra DNA sequences, and eliminates the need for extensive enzymatic treatments of the DNA such as restriction and ligation. The enzyme mix provided with the Seamless Cloning Master Mix recognizes and precisely assembles the DNA fragments sharing a 15-20base pair (bp) end homology that can be created by PCR-amplification.
The end result is a double-stranded fully sealed DNA molecule that can serve as template for PCR, RCA or a variety of other molecular biology applications, including direct transformation. Using the Seamless Cloning Master Mix, you can clone multiple DNA fragments into a single vector without subcloning, create modular expression vectors with interchangeable parts, construct seamless fusion proteins, delete and replace DNA segments, make internal protein fusions, swap tags on a gene, add UTRs to a cDNA, insert restriction sites, and more.
