Nonplastic_no-Care_1
Individual male brains of Nicrophorus vespilloides were dissected, snap-frozen liquid nitrogen after dissection, and stored at -80 C. We extracted RNA and genomic DNA (gDNA) simultaneously using Qiagen's AllPrep DNA/RNA Mini Kit (cat. # 80284; Hilden, Germany after homogenization with a Kontes handheld pestle (Kimble Chase, Rockwood, TN, USA). We prepared libraries for RNA-seq with a modified Smart-seq2 protocol (Picelli et al., 2014) using a target of 80 ng of total RNA per library and barcoded with Illumina TruSeq indexes. Libraries were SPRIed to select for fragments between 300-1000 bp and insert size was estimated with a Fragment Analyzer Automated CE System (Advanced Analytical, Ankeny, IA, USA). We sequenced 24 samples (six from each of the four behavioural states), assigned to one of two pools to evenly distribute samples based on experimental factors across the two lanes, with a 75bp single-end (SE) protocol using to Illuminas NextSeq500 with a High-Output flow cell at the University of Georgias Georgia Genomics Facility.