RNASeq of pea aphid line 218 in response to crowding
We put aphids at either crowded (12 aphids in a 3.5 cm diameter Petri dish) or solitary (1 aphid in a dish) treatments for 12 hours. After treatment, we removed embryos from adults and pooled eight carcasses per biological replicate. We extracted RNA from each sample using Trizol (Thermo Fisher) and isopropanol precipitation with an ethanol wash. We removed genomic DNA using Zymo DNAse and cleaned the RNA using the Zymo RNA Clean and Concentrator-5 kit under recommended protocols. RNA quality was verified on an Agilent 2100 Bioanalyzer. Libraries were prepared using the NEB Ultra RNA kit with NEBNEXT adaptors using 1ug of RNA as input under recommended protocols. After library construction, molarity of each library was checked with qPCR and equimolar amounts were multiplexed and loaded on the sequencer.