Synchrotron radiation with its high brilliance and tunable energy spectrum, is extensively employed in materials science, chemistry, biology and physics applications. Synchrotron radiation is passed through a set of optical components and required beam is transferred to a target sample. Since every dedicated experiment needs unique beam requirements for each sample, beamline components must be aligned specifically and exhibit specific properties. Current methods applied at beamlines rely on manual adjusting of each optical element which takes days even weeks. In this study, using genetic algorithms, the parameters of beamline components for achieving the smallest synchrotron beam spot size have been attempted for two different beamlines with focusing lenses and Kirkpatrick–Baez Mirrors. This study conclusively demonstrates that the adoption of genetic algorithms, integrating domain-specific knowledge, presents a notable temporal advantage over manual optimization. Moreover, it presents the viability of genetic algorithms as a proficient alternative to manual techniques at beamlines, thereby enhancing efficient use of beamtimes for end-users.