title: Generic sample from Papilio zelicaon
organism: Papilio zelicaon
description: Adult females were collected 2 populations of P. zelicaon,   near the latitudinal center of the species distribution (within 50 km of Medford, Oregon). Eggs   laid by these females were hatched under conditions characteristic of native habitats in a greenhouse and then moved to Conviron growth chambers at the University of Notre Dame. Multiple individuals of each larval instar were collected through the final instar before pupation. Individuals of the 3rd and 4th instars were exposed to a heat stress of 35 degrees for 60 minutes and a cold stress of 0 degrees for 120 minutes. Individuals in the  3rd and 5th instars of were exposed to a desiccation agent (silica gel) for   120 minutes. In addition, some of the collected larvae of P. zelicaon were fed Petrsolinum crispum and   others were fed Lomatium utriculatum. The former contains higher concentrations of linear furanocoumarins, a   defensive compound against herbivores, than the latter. After treatment, larvae were  frozen in liquid nitrogen and stored at -80 degrees C.    \textcolor{red}{Whole-body} RNA from these \textcolor{red}{frozen} individuals was extracted using an RNA Easy kit   (QIAGEN Inc.) \textcolor{red}{over a period of two months.} \textcolor{red}{Prior to library construction, pooling   was done by adjusting sample contributions to equimolar amounts of total RNA per individual.}  In total, 53 larvae of \textit{E. propertius}   and 61 larvae of \textit{P. zelicaon} were pooled prior to sequencing, representing a minimum of 11 and a maximum of 48 wild   mothers in \textit{E. propertius} and exactly 5 wild mothers for \textit{P. zelicaon}.