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The curriculum support materials used in an instructional process may influence assessment and achievement of learners. Curriculum support materials are both print and electronic in nature. One of the main electronic curriculum support materials used in schools is interactive digital content. In recent years, there has been rapid expansion in curriculum digital content access and utilization in Kenyan schools. However, little has been done to establish how utilization of curriculum digital content in the instructional process influences learners’ assessment and achievement. The study reported in this paper therefore investigated the influence of curriculum digital content utilisation on secondary school learners’ assessment and achievement in Biology. It was guided by two specific objectives including to: investigate the influence of the utilisation of Biology digital content on learners’ assessment and examine the influence of the utilisation of Biology digital content on learners’ achievement. A mixed methods approach that employed triangulation design was used in this study. Data was obtained from fifteen Economic Stimulus Programme-ICT phase 1 secondary schools in Nairobi County. Three instruments including Biology teacher questionnaire, learners’ focus group discussion guide and documents analysis sheet were used for data collection. The collected data was analyzed using both descriptive and inferential statistics and presented in form of notes, tables, and graphics. This study found out that most assessment aspects improved with curriculum digital content utilization. In addition, utilisation of curriculum digital content had a positive influence on the achievement of secondary school learners in Biology. The study recommended that all schools, and not only ESP-ICT schools, be provided with proper infrastructure to enable learners and teachers access and utilize curriculum digital content in the instructional process. This would enhance assessment modes and learners achievement in secondary schools in Kenya.