Purpose: To investigate the effects of refraction on retinal thickness measurements at different locations and layers in healthy eyes of Saudi participants. Methods: Thirty-six randomly selected adults aged 27.0 ± 5.7 years who attended a Riyadh hospital from 2016 to 2017 were categorized into three groups: non-myopic (spherical equivalent refraction [SER], +1.00 to –0.50 diopters [D]), low myopic (SER, – 0.75 to –3.00D), and moderate to high myopic (SER ≤ –3.25D). Full, inner, and outer retinal thicknesses were measured at nine locations by spectral-domain stratus optical coherence tomography (Optovue Inc., Fremont, CA, USA) and were compared according to refractive group and sex. Results: The mean SERs for the non-myopia, low myopia, and moderate to high myopia groups were 0.2 ± 0.6, –1.5 ± 0.5, and –7.5 ± 1.9 D, respectively. Refractive error, but not sex, had significant effects on the retinal layer thickness measurements at different locations (P < 0.05). The parafoveal and outer retinal layers were significantly thicker than the perifoveal and inner retina layers in all groups (P < 0.05). The full foveal thickness was higher and the full parafoveal and perifoveal regions were thinner in moderate to high myopic eyes than in the non-myopic eyes (P < 0.05), but were similar to those in the low myopic eyes (P > 0.05). The foveal thicknesses measured in the inner and outer layers of the retina were higher but the thicknesses measured at the inner and outer layers of the parafoveal and perifoveal regions were lower in moderate to high myopic eyes. Conclusion: There were regional differences in the retinal layer thicknesses of healthy Saudi eyes, which was dependent on the central refractions. This is important when interpreting retinal nerve fiber layer thicknesses in myopia and disease management in Saudi participants.