This study investigated the assessment of students’ performance in Senior School Certificate (SSC) multiple-choice test in biology. The survey research design was employed for the study. Sample for the study consisted of 1450 Senior Secondary Three (SSIII) students drawn from 20 randomly selected secondary schools in Ekiti State, Nigeria. The instruments used for the study were 2008 SSC National Examination Council (NECO) multiple choice Biology test paper. Findings from the study showed that significant different existed between students’ performance in NECO SSCE multiple choice test in biology on the basis of senatorial districts(F = 55.556, P<0.05), school location (t = 6.89, P<0.05),; gender(t = 5.03, P<0.05), and school type (F = 75.126, P<0.05). Based on the findings of this study, it was recommended that Government and other stakeholders in education should equip rural schools with facilities that are available in urban schools to improve students’ performance in Biology in rural schools.