RNAseq of Atta cephalotes: adult worker brains
Individual brains were dissected from workers that were snap-frozen only during 1100-1300 hr to minimize potential influences of circadian rhythms on gene expression. Individual samples belonging to different worker size groups and colonies were distributed across extraction batches, library batches, and sequence lanes to mitigate confounding batch effects. Total RNA was extracted from worker brains using a ThermoFisher PicoPure kit and a modified protocol for homogenization; three to 10 brains were pooled into a single microcentrifuge tube for each sample during the dissection process. Libraries were sequenced by Harvard BioPolymers using a combination of Illumina NextSeq and MiSeq with SE 75 reads. RNAseq unstranded libraries with mRNA poly-A selection were prepared by Harvard BioPolymers using a KAPA mRNA HyperPrep kit. mRNA sequence libraries were individually barcoded and multiplexed in equal proportions and all libraries were sequenced across four lanes.