BackgroundThe precise and efficient analysis of single‐cell transcriptome data provides powerful support for studying the diversity of cell functions at the single‐cell level. The most important and challenging steps are cell clustering and recognition of cell populations. While the precision of clustering and annotation are considered separately in most current studies, it is worth attempting to develop an extensive and flexible strategy to balance clustering accuracy and biological explanation comprehensively.MethodsThe cell marker‐based clustering strategy (cmCluster), which is a modified Louvain clustering method, aims to search the optimal clusters through genetic algorithm (GA) and grid search based on the cell type annotation results.ResultsBy applying cmCluster on a set of single‐cell transcriptome data, the results showed that it was beneficial for the recognition of cell populations and explanation of biological function even on the occasion of incomplete cell type information or multiple data resources. In addition, cmCluster also produced clear boundaries and appropriate subtypes with potential marker genes. The relevant code is available in GitHub website (huangyuwei301/cmCluster).ConclusionsWe speculate that cmCluster provides researchers effective screening strategies to improve the accuracy of subsequent biological analysis, reduce artificial bias, and facilitate the comparison and analysis of multiple studies.