RT Journal Article SR Electronic T1 Flexible open-source automation for robotic bioengineering JF bioRxiv FD Cold Spring Harbor Laboratory SP 2020.04.14.041368 DO 10.1101/2020.04.14.041368 A1 Chory, Emma J A1 Gretton, Dana W A1 DeBenedictis, Erika A A1 Esvelt, Kevin M YR 2020 UL http://biorxiv.org/content/early/2020/04/16/2020.04.14.041368.abstract AB Liquid handling robots have become a biotechnology staple1,2, allowing laborious or repetitive protocols to be executed in high-throughput. However, software narrowly designed to automate traditional hand-pipetting protocols often struggles to harness the full capabilities of robotic manipulation. Here we present Pyhamilton, an open-source Python package that eliminates these constraints, enabling experiments that could never be done by hand. We used Pyhamilton to double the speed of automated bacterial assays over current software and execute complex pipetting patterns to simulate population dynamics. Next, we incorporated feedback-control to maintain hundreds of remotely monitored bacterial cultures in log-phase growth without user intervention. Finally, we applied these capabilities to comprehensively optimize bioreactor protein production by maintaining and monitoring fluorescent protein expression of nearly 500 different continuous cultures to explore the carbon, nitrogen, and phosphorus fitness landscape. Our results demonstrate Pyhamilton’s empowerment of existing hardware to new applications ranging from biomanufacturing to fundamental biology.Competing Interest StatementThe authors have declared no competing interest.