Recombination, linkage and crossover of chromosomes are the key and difficult points in genetics teaching. For students’ comprehensively understanding these concepts in teaching, we designed a new genetic experiment named ‘Gene mapping using DNA molecular markers’ based on the principle of the map-based cloning method, using our previous studies on cloning of a rice lesion-mimic phenotype regulating gene SPOTTED LEAF 5 (SPL5) as an example. In this experiment, students are instructed to perform the linkage analysis, genetically map the spl5 mutation and align a genetic map by using an F₂ population generated by a cross between spl5 and wild-type plants and polymorphic molecu- lar markers. This teaching experiment can not only effectively promote students' understanding of the three Laws of genetics, but also be helpful to broaden their vision and improve their ability to solve problems and work in a team.