TNT Cloning System

Organization: Oak Ridge National Laboratory
Year: 2018

The TNT Cloning System is designed for flexible DNA assembly from universal libraries to generate multigene constructs (DNA synthesis). This cloning system adopts a predefined three-nucleotide (TNT) signature and a buffer system for a quick one-pot reaction. The allinclusive and ultra-flexible TNT Cloning System assembles functional constructs from a universal library that automatically maintains open reading frames and does not require linkers, adaptors, sequence homology, amplification, or mutation of DNA fragments to work properly. The enhanced buffer formulation provides the capability for testing and sharing complex multigene circuitry composed from a variety of DNA fragments. The system has also been engineered to shuttle between different biological systems. The easily transferable platform can be used to assemble a gene from a randomized library in vectors ready for transformation in a variety of hosts including bacteria, yeast, plant, and mammalian cells.

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