DNA assembly for plant biology: techniques and tools

•Plant biotechnology and synthetic biology require the assembly of complex DNA molecules.•Several one-step DNA assembly methods have been developed.•These either utilize restriction endonucleases or overlapping, linear DNA fragments. As the speed and accuracy of genome sequencing improves, there are ever-increasing resources available for the design and construction of synthetic DNA parts. These can be used to engineer plant genomes to produce new functions or to elucidate the function of endogenous sequences. Until recently the assembly of amplified or cloned sequences into large and complex designs was a limiting step in plant synthetic biology and biotechnology. A number of new methods for assembling DNA molecules have been developed in the last few years, several of which have been applied to the production of molecules used to modify plant genomes.