Gender factor can have a significant impact on the organization or even an individual with certain gender characteristics can be different from organization to organization, the degree of acceptance of the models and assumptions underlying the organization. The purpose of this study was to investigate the relationship between gender and culture at the National Library of the Islamic Republic. This study applied a descriptive- survey. For collecting the survey data, standard questionnaire used. Denison with 60 questions and four sections were collected. Statistical analysis, both descriptive and inferential statistics using SPSS 20 software has been made. The results showed that the average overall score of "corporate culture based on the National Library of Dennison" Less than average level is. Most points after engaging in work (participatory) (mean 01.3) and lowest points in the mission rocker (Mission) (mean 83.2) have been obtained in the field theory has been achieved. The results showed that among the four dimensions of organizational culture that includes gender and conflict at work (participatory), adaptability (stability and integrity), adaptability (flexibility) and taking the mission (mission) are not statistically significant and researcher's claim was not confirmed and the observed difference between the mean scores based on gender, according to the twelve indicators of organizational culture, empowerment, team building, development potential, intrinsic value, consistency and coherence, flexibility, customer orientation, organizational learning and landscape variables, gender was not significant, but Indicators such agreements, strategic direction and goals and this difference was significant.