The TNT Cloning System allows for speedy multi-gene DNA synthesis and assembly from universal gene libraries. The system is all-inclusive, capable of handling large DNA constructs at speeds up to 80% faster than other assembly systems.
It adopts a pre-defined three-nucleotide signature—TNT—and a buffer system for quick one-pot reactions. Enzymes create specific signatures that allow three elements to be combined at once per round of assembly. This universally exchangeable system is transferable across hosts including bacteria, yeast, plant and mammalian cells.
This system is unique, as it does not require adaptors, sequence homology, amplification or fragment mutation to construct DNA. The technology allows for greater access among research communities and advances cheap and environmentally friendly bio-manufacturing efforts.
The development team, led by ORNL’s Gerald Tuskan, included Xiaohan Yang of ORNL and Henrique De Paoli, formerly of ORNL and the University of Tennessee, currently of Lawrence Berkeley National Laboratory.
The TNT Cloning System was funded by the ORNL Technology Innovation Program and the DOE Office of Science’s Genomic Science Program.