AbstractHemoglobinopathies are one of the major problems in Pakistan. A retrospective analysis of blood samples of 2731 patients from 2010 to 2014 was done at National Institute of Blood Disease & Bone Marrow Transplantation for the workup of anemia or other blood-related disorders. Whole blood samples in EDTA were collected; complete blood counts with peripheral smears were prepared. Hemoglobin (Hb) electrophoresis on Genio was performed at alkaline pH. Samples showing borderline results were further tested by high-performance liquid chromatography or for specific mutation analysis by ARMS-PCR. Out of total 2731, 935 (34.2%) patients had hemoglobinopathies. Out of these 935 patients who had hemoglobinopathies, beta thalassemia minor 51.8%, beta thalassemia major 24.1%, HbD trait 6.7, sickle/beta thalassemia 4.5%, sickle cell disease 3.9%, HbE trait 1.9%, and sickle cell trait 1.7% were most common hemoglobinopathies. Less prevalent were delta/beta thalassemia, HbE homozygous, HbD homozygous, and HbH disease.