The extent to which Biology is failed by learners is high yet it is important in a medical and other scientific field. This level of failing has been closely associated with the teaching methods applied by the biology teachers. The main objective of this study was to assess how teaching methods affect students’ performance of biology in Soroti Secondary School, Soroti district. The study specifically determined the relationship between teaching methods and students learning of Biology, identified the resources necessary for the proper teaching of Biology and ascertained the challenges faced by the students while learning biology in Soroti Secondary School. The study employed a descriptive cross-sectional study design using a quantitative approach for data collection with questionnaires and interview guides as the research tools while applying questionnaire and interviews as data collection methods to extract primary data from 81 respondents. The results indicated that (46.9%) of the respondents used explanations as their Biology teaching method, (59.3%) affirmed that their learners understood Biology well using the methods applied by them to teach, and (67.9%) observed that their learners performed highly in Biology due to the methods of teaching they used. Additionally, (52%) said that biological laboratories were adequate, (84%) asserted that Biological supplementary books were inadequate, (60%) reported that teaching guides were underutilized and (56%) said that Biology teachers were inadequate. Furthermore, (67.9%) reported that Biology as a subject was feared, (75%) said that they were not able to access all the resources for Biology, (54.3%) described the relationship between the teachers and the learners to be poor and (57%) reported that there was absenteeism of learners from school. The study therefore concluded that there was a very strong positive relationship between teaching methods and students’ performance of Biology at r 0.708 and at level of significance 0.05, this implied that if the proper teaching methods for Biology were used, the level at which students would pass Biology would be high and if improper teaching methods of Biology were used, the students would fail Biology.