We propose a modified and updated protocol to obtain mitotic chromosomes from the regenerated tissue of Pelophylax tadpole tail tips. Chromosomal preparations from regenerated tissue results in high-quality and clean slides suitable for further staining and study. Tadpoles remain alive, undergo minimum suffering, and can be grown to adulthood for further investigation. The method could be used for other groups of Anura and modified for other species with the ability to regenerate their tissues.