RNA-Seq of Eucera pruinosa: pooled adult and larval tissue
RNA was extracted from four individuals: two adult males (whole heads), one adult female (head and abdomen), and one larva of unknown sex (head, middle section, and tail). Extractions were performed with a Qiagen RNeasy kit following the manufacturer's recommendations, including DNA degradation with DNase I and a final elution in 50 l of nucleic acid-free water. Concentration of each sample was measured with a Qubit fluorometer, using the high-sensitivity RNA reagent. Quality was checked with an Agilent TapeStation.  RNA from all eight extractions was pooled in equal concentrations and sent to Novogene (Davis, CA) for library preparation and sequencing. A single mRNA library was prepared using poly-A enrichment via oligo(dT) beads, and a custom second-strand synthesis buffer (Illumina). Library insert size was checked on an Agilent 2100 and concentration was quantified with qPCR. Sequencing was performed on an Illumina NovaSeq 6000 machine.