Objective: To compare the frequency of post-operative wound infection with single layer versus multiple layer closure of wound in patients undergoing major surgeries. Study Design: Quasi-experimental Study. Place and Duration of Study: Department of Surgery, Combined Military Hospital, Rawalpindi Pakistan, from Aug 2018 toAug 2019. Methodology: After getting written consent and considering the inclusion criteria 150 patients were chosen for this study. They were indiscriminately separated in two groups by using chit method. In Group A, patients underwent surgery with single layer closure. In Group B, patients underwent surgery with multiple layer closure. Most of surgeries were done under general anesthesia. Then post-operative wound infection was noted in the patients of both groups by using proforma. SPSS version 20 was used for all the collected data analysis. Results: In this study mean age of the patients in Single Layer Group was 48.17±9.07years and in Double Layer Group was 51.55±18.72 years. Male to female ratio was 1:1.03. The most common cause of surgery was colostomy closure found in 34(22.67%) patients. Wound infection noted in 19(12.67%) patients. In Single Layer Group patients the wound infection was found in 4(5.3%) patients whereas in Double Layer Group the wound infection was noted in 15(20%) patients (p-value=0.007). Conclusion: Single layer closure is safe technique which showed significantly less post-op wound infection than to double layer closure of skin in patients undergoing major surgeries.