This study investigated the prevalence of previously reported mutations in the Otoferlin gene, implicated in the non-syndromic autosomal recessive deafness (NSARD) disorder known as DFNB9. 63 unrelated Omani patient samples, previously diagnosed with NSARD, were chosen for this study. An equal number of healthy Omani individuals were chosen as controls. The mutations screened were: (1) c. IVS24 2856 +1 G>A, (2) c.836 C >T(.Arg 237 Ter), (3) c.2416 T>A (p. Tyr 1496 Ter) and (4) c.2485 C> T (p.Glu 829 Ter). The mutations selected for screening had been reported in Spanish and Middle eastern populations such as the Lebanese and Emiratis. PCR-RFLP and Direct DNA sequencing were used for mutation detection. However, none of the screened mutations were detected in any of the 63 Omani patients tested. Two new, unreported polymorphisms were noted in patient samples, of which, one was a point mutation, also detected in some control samples, where a single nucleotide change G>A at position 499 did not result in any change in amino acid (Thr 124 Thr). The other polymorphism, which was also a single nucleotide change at 494 (G>A), resulted in the replacement of a Glycine by a Serine at codon 123 (Gly 123 Ser). This mutation occurred close to the predicted C2A domain of otoferlin. Hence, a thorough analysis was carried out using bioinformatics analytical tools and software to evaluate the significance of this mutation in the etiology of deafness. In the course of these analyses, a putative and partial 3-D model of Otoferlin was produced on the basis of Homology/Comparative Modeling principles incorporated in the SWIS MODEL program available on the Internet. The partial. 3-D model enabled a simulation study of the possible effects of the c.494 G>A (p. Gly 123 Şer) mutation detected, on the tertiary structure characteristics. It was concluded that this mutation was silent using primary, secondary and tertiary structure bioinformatics analyses techniques. Thus the new c.494 G>A (p. Gly 123 Ser) mutation discovered in this study does not contribute to NSARD.