Background and Aims: Diabetic ulcers are a major cause of morbidity and mortality. Identifying and isolating the causative microbe helps in control of infection. The aim of the study is to compare swab culture and tissue culture for the routine identification of pathogens in diabetic wound infection. Materials and Methods: This was a prospective observational study conducted in patients admitted at Surgery ward with diabetic ulcers in the months of June and July 2019. The patients were classified according to the PEDIS grading. Wound swab and tissue specimen were obtained by Levine technique and punch biopsy respectively and processed in the microbiology lab to identify the causative pathogen in the wound. Results: Of the 72 patients observed, males were affected more (72.2%) and many had grade 3 ulcers (86.1%). While Klebsiella was the predominant organism (42.9%) in tissue isolates, Pseudomonas was predominant in swab isolates (41.2%). Swab cultures were more effective in detecting microbes (40%) in grade 2 wounds, while tissue cultures were more effective in grade 3 wounds (70%). The mean of swab culture and tissue culture were 4.25 and 4.00 respectively in student’s paired t–test. Conclusion: Swab culture was more reliable for detecting microbes in grade 2 diabetic ulcers while tissue culture along with swab culture are needed for ulcers of grade 3 and above to aid in appropriate wound healing