Fig. 5

Comparison of gene expression analysis in two biological replicates by RT-qPCR, performed using automated and manual procedures. The graphs report the gene expression, represented as mRNA relative quantity (mRNA RQ), of Ciona juveniles exposed to Pam2CSK4 (1 μg/ml) or zymosan (100 μg/ml), for 30 min, 2 h, and 4 h. (A), “Biological replicate 1”, resulting in a more effective response to the microbial stimuli treatment, reflects a clearer comparison between the two procedures. In contrast (B), less responsive animals (“biological replicate 2”) display higher variability in gene expression detection