Biotechnology has been widely applied in various products throughout the 21st century. Malaysia selected the biotechnology sector as one of the key strategic technologies that would enable Malaysia to transform into a fully developed nation by the year 2020. However, to date, there has been very little research on the level of biotechnology literacy amongst school students in Malaysia and the aim of this study is to determine the presence of biotechnology literacy, based on knowledge, perception, and attitude. Besides, this study also determines the factors affecting the students’ attitude towards biotechnology. This study is a quantitative study involving 292 secondary school students from science and non-science classes. The findings showed that overall students have what we describe as a medium level of knowledge, perception, and attitude towards biotechnology. It is anticipated that this pattern of findings can help teachers, the Ministry of Education and the media to strategically plan ahead so that improvements in the national curriculum can be proposed and implemented.