In this study, the large scale forward genetic screening had been done in Thor lab, in order to distinguish regulator genes and mechanisms behind stem cell development process by utilizing an accurate single cell marker (FMRFa). In brief, 7 lines of EMS random mutagenized flies with TV-GFP reporter insertion, were homozygosed for the mutant chromosome. They had shown loss of FMRFa and alerte Eya Expression at 18 AEL ST.

Furthermore, those mutants were deletion based mapped for lethality and phenontyped to identify the underlying neurodevelopmental regulatory genes. This step was followed by GFP expression analysis, dissections of the CNS, antibody staining, microscopy and confocal microscopy. Finally, molecular mapping (PCR and gene sequencing) reaveled some candidate mutations for the phenotype of interest in the identified gene.