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Microstructure strongly influences the mechanical properties of cast iron. By inoculating the melt with proper inoculants, foreign substrates are brought into the melt and eventually the graphite can crystallize on them. The elements and substrates that really play a role for nucleation are yet unknown. Until now there is very little knowledge about the fundamentals of nucleation, such as composition and morphology of nuclei[1,2]. In this work we utilized EN-GJL-200 as a base material and examined several produced specimens. The specimens were cast with and without inoculants and quenched at different solidification states. Specimens were also examined with a high and low oxygen concentration, but the results showed that different oxygen contents have no influence on the nucleation in cast iron melts. Our research was focused on the microscopic examination and phase-field simulations. For studying the samples we applied different analytical methods, where SEM-EDS, -WDS were proved to be most effective. The simulations were conducted by using the software MICRESS®[3], which is based on a multiphase-field model and has been coupled directly to the TCFE3 thermodynamic database from TCAB. On the basis of the experimental investigations a nucleation mechanism is proposed, which claims MnS precipitates as the preferred site for graphite nucleation. This theory is supported by the results of the phase-field simulations.