This Small Business Innovation Research (SBIR) Phase I proposal requests funding to develop a novel technology for the controlled non-templated enzymatic synthesis of single strand DNA molecules for use in molecular, systems, and synthetic biology applications. When implemented, this technology represents the most significant change to the way that oligodeoxynucleotides and polydeoxynucleotides have been produced since the introduction of the phosphoramidite mediated solid support method in 1981. The new process will relieve the bottle necks in molecular and synthetic biology workflows by reducing costs, increasing the lengths of DNA directly synthesized by 10x to 50x, reducing turnaround times from weeks and months to days, and eliminating production of tons of toxic wastes. A unique enzyme will be used that catalyzes the rapid polymerization of deoxynucleotides without need of a template molecule. The project will create novel analogs that are suitable enzymatic substrates and allow the stepwise addition of nucleotides mimicking the gold standard chemical synthesis method but in a simpler and aqueous enzymatic process.<br/><br/>The broader impact/commercial potential of this project, if successful, will be to create a next generation approach to DNA synthesis. Such capabilities will not only enhance current research, but will enable evolving multidisciplinary research areas such as biomedicine, computer science, nano-optoelectronics, and bionanotechnology. Research utilizing DNA nanostructures for drug delivery and single cell in-vivo analysis is evolving. DNA is being used as an information storage medium and for computing itself. Engineers are investigating gene circuits as biological analogs of electronic components such as oscillators and transistors. This most interesting, complex and biologically important molecule surely has many applications that cannot yet be imagined. Just as the first generation DNA synthesis technology enabled a new era of biotechnology 25 years ago, the capabilities presented here will enable a new generation of biological applications and potentially provide for society's future needs.