title: RNA-seq of Timema tissues
design: Prior to RNA extraction, animals were fed with artificial medium for two days to avoid contamination with gut content and then frozen at -80ÁC. Total RNA was then extracted. This was done by first freezing the individuals in liquid nitrogen, followed by addition of Trizol and mechanical bead crushing. The homogenized tissue was then treated with Chloroform and Ethanol and the aqueous layer transferred to RNeasy MinElute Columns (Qiagen). Following the RNeasy Mini Kit protocol, potential DNA in the sample was digested, RNA eluted into water and stored at -80ÁC