Fig. 2

The robot-automated method enables precise gene expression analysis by RT-qPCR. a Gene panel selected to evaluate early gametocytogenesis-related gene (GRG) expression dynamics. The basal expression of each gene in the panel was evaluated for ring- and trophozoite-stage cultures, which continuously commit and generate a basal level of gametocytes expressing these genes. Technical repeatability was evaluated by the standard deviation value (error bars) of the Ct in three independent biological repeats (n = 3). b SPUD polymerase inhibition assay for cDNA synthesized by robot-automated methods from ring- and trophozoite-stage parasites. Artificial SPUD template at a 10^− 5 nM concentration was used as a non-inhibition control, and phenol was used as a positive inhibition control. No Amp. = no amplification (Ct ≥ 40). a–b Mean of three biological replicates (n = 3) with three technical replicates each. Error bars represent SD. A two-way ANOVA with Šidák’s multiple comparisons test was performed for Panel b. ns = p ≥ 0.05 (non-significant)