Developmental time course of F48E3.8 and C54G7.3 gene expression in germline-ablated <i>C. elegans</i> using RT-PCR.

<p>Synchronous non-permissive (germline-ablated) worms of this strain were grown on solid media and used to collect RNA from various time-points following L1 arrest at hatching and continuing into adulthood. First strand cDNA (+) was generated from each RNA extract (including No RT controls, –) and used as a template for PCR to analyze expression of the genes F48E3.8, C54G7.3, the eggshell chitin synthase <i>chs-1</i> and the pharyngeal chitin synthase <i>chs-2</i>. The gene <i>ama-1</i>, encoding the large subunit of RNA polII, was used as a positive control (as done by Johnstone and Barry 1996 and Veronico <i>et. al.</i> 2001) although we note that expression of the gene is not consistent as previously described. Ablation of the germline was confirmed by the attenuation in expression of <i>chs-1</i>. The transition to adult stages was confirmed by the expression of <i>col-19</i>, an adult specific collagen gene. All primers and PCR conditions are listed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0040426#pone.0040426.s002" target="_blank">Table S2</a>.