Drosophila PGC transcriptome

To understand transcriptomic dynamics of PGC (primordial germ cells; pole cell) of fruit fly, we used fluorescence-activated cell sorting (FACS) to isolate PGC from the embryos carrying the transgene, EGFP-vasa, which expresses GFP specifically and continuously in the germline throughout the life cycle. The PGCs collected at 11 time points during the embryogenesis were subjected to microarray analyses. RNA from FACS-sorted PGCs collected at 11 different time points were hybridized to a microrray after Cy3/Cy5 labeling. Four indepent experiments (biological replicates) were conducted for each time point. A pooled total RNA purified from whole embryos at early to late stage was used as a common reference in all hybridizations.