The complex abilities of pitch perception and perceiving discriminating pitches are vital to the music perception phenomenon. Comprehensive studies have investigated the influence of intervening silence in various dimensions and have indicated its effects on human auditory ability. The present study provides additional information on the pitch discrimination ability, with a focus on how silence between stimuli affects our perception. We neutralized various effective factors and assessed the effect of perceivable durations of silences (in the range of 500-2000 ms) on pitch discrimination ability in musicians; distractive factors were reduced to refine the effect of intervening silence, and assess pitch discrimination accuracy in varying silent inter-stimulus interval durations precisely. By using factors such as pure stimuli, moderate frequencies, suitable durations, and favorable intensities, the efficiency of the experiment was optimized. Varying degrees of pitch differentials (5, 10, 25, and 50 cents) were used to estimate the effect of silence on pitch discrimination. In this experiment, participants included 36 female and 65 male undergraduate and graduate music students from Tehran University of Art and University of Tehran (mean age=23. 31 years, SD=4. 46). They had passed ear-training courses, and played a melodic instrument for a minimum mean (SD) time of 9. 15 (3. 42) years. The difference limen for frequency was es-timated for all participants (mean± SD= 8. 2± 2. 88 cents). Since the results of multiple test attempts on one person should be considered related or dependent, Friedman test was used to compare the answers of each participant across different silent ISI durations. Moreover, Fisher’ s exact test was used to correct all analyses. To assess the influence of auditory sensitivity and musical experience in our experiment, and to find any correlation between these factors and the effect of silence intervention, par-ticipants were categorized into two groups: one based on their baseline mean DLF (8. 2≤ and >8. 2) and another based on the mean years of their musical instruction (≥ 9. 15 years or <9. 15 years). Friedman tests were again used to examine the difference between the answers of these groups. To perform a detailed analysis, logistic regression was used to assess the relationship of each duration of silence to the correctness of pitch discrimination (0 ms silent ISI as reference). All analyses were done using IBM SPSS for Windows version 19 with a two-tailed P value < 0. 05 being considered significant. The results showed that improvement in pitch discrimination ability was significant when the duration of silence occurred between stimuli in most of discriminations (p<0. 05). In addition, the auditory sensitivity and musical experience factors had no significant effect on this phenomenon. This effect is defined by the mechanical functions of the inner ear or procedures of tonal memory in the auditory process. Interestingly, the examined durations of silences (500, 1000, and 2000 ms) can conveniently be calculated by every common metronome. Hence, our achievement can be applicable to music educators to set better conditions reinforcing the pitch discrimination ability in the practice procedures or instruction processes. In addition, this information advantageous for investigators as an executable implement to design specific experiments in pitch discrimination assessments, and any other psycho acoustical researches.