Musca domestica Embryo
For RNA sequencing, we selected four early stages of M. domestica embryogenesis and collected 100 embryos from each one. RNAs were extracted using TRIzol (Lifetechnologies) according to manufacturer instructions. Integrity of RNA samples was examinated by electrophoresis on denaturing agarose gel. Analysis of absorbance using an Infinite 200 PRO Nanoquant (TECAN) spectrometer was used to quantify RNA mass and discard those samples with 260:280 ratio under 2, to avoid protein containation. Each sample was Ambion Turbo DNAse and purified using the RNA Clean Up protocol from the RNAeasy Mini Kit (Quiagen). Messenger RNA was isolated from each sample using the Micropoly(A)Purist Kit from Life Technologies, according to manufacturer instructions. An equal mRNA mass of each sample were pooled and used for sequencing.