Temp = 20C
Specimens were transported live to the laboratory within 24h of collection and held under common garden conditions at 11C for two weeks before being transferred to tanks for different temperature conditions.  Tank temperatures were ramped over 4 days to temperatures ranging from 15.5C to 28.6C at which time they were held for 24 hours prior to sampling.  Transcriptomic (RNA-seq) analyses were based on n=32 specimens, with n=8 per temperature treatment . Thorax tissue was homogenized under liquid N2 using a TissueLyser II (Qiagen 85300) for 5-10 sec and RNA was extracted using the manufacturers polysaccharide/proteoglycan removal modified protocol, using BCP and High Salt Precipitation Solution (MRC). Purified RNA was assessed for quantity and quality using a Bioanalyzer (Agilent) and only samples with little to no degradation and adequate concentration were used in subsequent steps.  For each exposure temperature, equal amounts of total RNA from each individual were used to make n=5 independent pooled RNA samples.  Those n=5 pooled RNA samples were used to make RNAseq libraries following the Illumina Tru-Seq RNA v2 kit