There are various causes of human male infertility and various genes might be involved. Gene under study for present research work was GNRH1 (Gonadotropin Releasing Hormone 1). For this study 40 samples of males with abnormal semen analysis report were collected from different diagnostic centers with informed consent. Then DNA was extracted. Primers were designed for GNRH1 gene and amplified through PCR. Sequencing was outsourced and bioinformatics tools were applied to analyze the sequenced data. There were 14 variations at different positions identified in this study in GNRH1 gene. Variations observed at position c.1325+40 and c.1325+181 mentioned in literature as rs2709608 but predicted that it has no significant effect on protein, whereas rs1453947741 have not been cited in any research article till date which is associated with deletion of A at position c.1325-144. It is potential target site and shows significant change affecting splicing. No data has been found which state the disease causing variations of diagnosed positions in my study. The study concluded that GNRH1 gene has certain variations and splicing might have effect on intronic variations which result in diseased condition and might have role in association with human male infertility.