Purpose. To evaluate the relationship between retinal nerve fiber layer (RNFL) thickness measured by optical coherence tomography (OCT) and the axial length/refractive error of the eye. methods. A total of 115 eyes of 115 healthy subjects, comprising 75 eyes with high myopia (spherical equivalent [SE] < −6.0 D) and 40 eyes with low to moderate myopia (SE between −6.0 D and −0.5D), were analyzed in this cross-sectional study. Total average and mean clock hour RNFL thicknesses were measured by OCT and compared between the two myopia groups. Associations between RNFL measurements and axial length and spherical equivalent were evaluated by linear regression analysis. results. The RNFL measurements were significantly lower in the high myopia group compared with those of the low-to-moderate myopia group at 12, 1, and 7 o’clock (right eye orientation). Apart from the temporal clock hours, significant correlations were evident between RNFL measurements and the axial length and spherical equivalent. The average RNFL thickness decreased with increasing axial length (r = −0.314, P = 0.001) and negative refractive power (r = 0.291, P = 0.002). A significant proportion of myopic eyes were classified as outside normal limits, with reference to the normative database. The most frequently abnormal sector was at 2 o’clock, where 16.5% of myopic eyes were outside normal limits. conclusions. RNFL measurements vary with the axial length/refractive error of the eye. Analysis of RNFL thickness in the evaluation of glaucoma should always be interpreted with reference to the refractive status. Although the normative database provided by OCT has been helpful in identifying ocular diseases involving the RNFL, it may not be reliable in the analysis of myopic eyes.