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Objectives This study investigated occupational exposure to radiofrequency electromagnetic fields (RF EMF) using two job-exposure matrices (JEMs) and risk of glioma.Design Population-based family case–control study.Setting Cases were recruited from participating hospitals in the Australian states of New South Wales, Queensland, Tasmania, Western Australia and Victoria between January 2013 and November 2017.Participants The study population consisted of 467 cases of glioma and 367 family controls recruited for the Australian Genomics and Clinical Outcomes of Glioma case–control study between 2013 and 2017. Participants completed questionnaires on demographic and other information, including a detailed occupational history.Exposures Exposure to RF EMF was estimated using both the multicountry case–control study INTEROCC JEM and the Canadian JEM (CANJEM).Primary outcome measures ORs and 95% CIs were calculated from logistic regression models adjusted for relatedness between cases and controls, sex, age, ethnicity, education level, smoking status and alcohol consumption.Results There was no statistically significant positive association overall for risk of glioma when applying either JEM. For the highest compared with the lowest quartile of lifetime exposure, results using the INTEROCC JEM showed an OR of 0.74 (95% CI 0.47 to 1.15) for electric fields and 0.92 (95% CI 0.58 to 1.45) for magnetic fields, while the CANJEM showed an OR of 0.85 (95% CI 0.54 to 1.32). We also did not observe associations when applying different assumptions regarding latency or time windows or with glioma grade.Conclusion Overall, this study found no evidence of an association between occupational RF EMF exposure and glioma. Future research should focus on refining occupational RF EMF exposure assessment.