Transcriptome analysis of the predatory mite, Phytoseiulus persimilis
Mites were collected and homogenized in Tri-reagent (Sigma-Aldrich) for extraction of total RNA. The oligotex kit was used to isolate mRNA, which was precipitated with sodium acetate and ethanol. A cDNA first strand synthesis was set with this mRNA sample, using the smart IV oligo and a 3'modified primer. The number of cycles for the second strand synthesis was optimized (25 cycles) and then 5 different PCR reactions were set to obtain enough cDNA material. Five micro grams were submitted to the Genome Science Laboratory at NCSU for pyrosequencing with the 454 technology.