The Cambrian explosion is a grand challenge to science today and involves multidisciplinary study. This event is generally believed as a result of genetic innovations, environmental factors and ecological interactions, even though there are many conflicts on nature and timing of metazoan origins. The crux of the matter is that an entire roadmap of the evolution is missing to discern the biological complexity transition and to evaluate the critical role of the Cambrian explosion in the overall evolutionary context. Here we calculate the time of the Cambrian explosion by an innovative and accurate “C-value clock”; our result (560 million years ago) quite fits the fossil records. We clarify that the intrinsic reason of genome evolution determined the Cambrian explosion. A general formula for evaluating genome size of different species has been found, by which major questions of the C-value enigma can be solved and the genome size evolution can be illustrated. The Cambrian explosion is essentially a major transition of biological complexity, which corresponds to a turning point in genome size evolution. The observed maximum prokaryotic complexity is just a relic of the Cambrian explosion and it is supervised by the maximum information storage capability in the observed universe. Our results open a new prospect of studying metazoan origins and molecular evolution.