Intervention(s): Comparison of sperm DNA fragmentation levels (DFLs) using fresh, snap-frozen and air-dried semen, with air-dried samples stored at different temperatures and time periods to assess DNA stability.Objective: To develop a simple, convenient, and stable storage method for semen before DNA fragmentation testing.Design: Experimental cross-sectional study.Setting: Fertility clinic.Patient(s): 164 male partners of infertile couples.Main Outcome Measure(s): DFL determined by Halosperm G2 kit.Result(s): Results are expressed as mean ± standard error of the mean. The DFLs from fresh and air-dried semen gave comparable results (1.08% ± 0.65%), and from snap-frozen and fresh samples a statistically significant difference (5.5% ± 1.09%). Air-dried semen stored at room temperature for 7 days had a statistically significantly higher DFL compared with semen stored overnight (46.29% ± 9.12%). Samples stored at 4°C for 7 days or 1 day showed no statistically significant difference (0.83% ± 0.82%). DFLs from samples stored for either 1 or 30 days at 4°C showed a statistically significant difference (19.59% ± 5.72%); those stored at -22°C showed no statistically significant difference (0.68% ± 0.53%).Conclusion(s): Air-drying semen is a simple and stable storage method for up to 1 month at -22°C before DNA fragmentation testing.