Background: Infertility is considered one of the main public health issues, as it affects about 15% of the couples of reproductive age. The male factor is involved in 40% - 50% of infertility cases. It is difficult to assess accurately the overall magnitude of the contribution of genetics to infertility as most, if not all, conditions are likely to have a genetic component. The genetic causes of infertility are varied and include chromosomal abnormalities, single gene disorders and phenotypes with multifactorial inheritance. Some genetic factors influence males specifically, whereas others affect both males and females. Frequently, however, in the clinic no clear cause for the observed infertility could be diagnosed which at least in part, it reflects our still poor understanding of the basic mechanisms that regulating and controlling the genetic networks of male infertility. Objective: To study the molecular identification of Azoospermic infertile patients by the gene sequencing of NR5A1. Subjects, Materials and Methods: A study carried out during the period from November 2014 to June 2015. Fifty specimens were collected from thirty azoospermic patients and twenty normal healthy subjects (normozoospermic subjects) as control their age ranged between 23 and 48 years old, the seminal fluid of cases indicated that liquefaction time, color and viscosity equal in normozoospermic subjects (control group) and patient group while there is significant difference P≥0.05 in sperm count reach to more than 20 million and motility reach to more than > 25% in control subjects while there is no sperm count found in azoospermic patients. Then blood was collected for hormonal assay, in azoospermic patients the results revealed a gradual decrease in serum testosterone levels with a concomitant increase in serum follicular stimulating hormone (FSH) level when compared with control subjects . NR5A1 gene were investigated in 30 samples of extracted DNA from azoospermic patients by using polymerase chain reaction (PCR) method it can directly detect the NR5A1 gene content after it had been molecularly identified then were sent successfully for sequencing analysis ,this study examined the presence of two transversion and one transition mutation of NR5A1 gene in azoospermic patients compared with control subjects, there’s no mutation in the same gene when compared with gene bank.