Developmental time course of gene expression in Anopheles gambiae embryo

In order to examine the gene expression in the course of mosquito embryogenesis, microarray assays were performed on staged A. gambiae embryos, from fertilization to 52 hours of development (which is close to hatching at ~50 hours post-fertilization). RNA was extracted from staged embryos roughly every three hours after fertilization, and then hybridized to the A. gambiae transcriptome microarray. Keywords: time-course Total RNA from unfixed staged eggs at sequential (2h, 4h, 6h, 7h, 8h, 10h, 13h, 16h, 19h, 22h, 25h, 28.5h, 31h, 34h, 37h, 40h, 43h, 46h, 49h, 52h) time points was purified initially by Trizol (Invitrogen) and then further by RNAeasy (Qiagen) kit. Biotin-labeled probes were synthesized using regular labeling kit (Affymetrix) and hybridized to Plasmodium-Anopheles expression arrays (Affymetrix). All time points were sampled with three independent biological replicates, except for time points 49h and 52h, which were sampled once. RMA-express software was used for processing of raw expression data and estimation of log base 2 expression values.