EisaIllumina
RNA was extracted separately from six adult tissue (four wings; three heads; four antennae, legs and mouth parts; thorax; abdomen 1-3, abdomen 4-6), and five tissue parts from early ommochrome stage of the pupae (head and mouth parts; wings, antennae and legs; thorax; abdomen 1-3, abdomen 4-6). Each tissue was frozen in liquid nitrogen and quickly homogenized in 500 l Trizol, adding the remaining 500 l Trizol at the end of the homogenization. Phase separation was performed adding 200 l of cold chloroform. The upper phase was then transfer to RNeasy Mini spin column and processed with Qiagen RNeasy Mini Prep Kit (Qiagen, Valencia, CA) and DNAse purification using the Turbo DNA-free kit (Life Technologies, Carlsbad, CA, USA) following the manufacturer's instructions. All the extractions were finally pooled keeping the same final RNA concentration from all samples.