RNA-Seq of cell-free hemolymph collected at 4 PM: adult male E. scolopes
Pooled cell-free hemolymph from 2 adults was used for RNA purification at each time point. Total extracted RNA was purified using mirVana PARIS kit (Invitrogen) and followed by treatment with DNAse I (Thermo Fisher Scientific). The RNA concentration was determined with a Qubit RNA BR assay kit (Invitrogen). The small RNA libraries were constructed using the TruSeq Small RNA Sample Preparation kit according to the manufacturers instructions. The quality of RNA libraries was assessed with Agilent 2100 Bioanalyzer System. The single-end 50 cycle sequencing was performed using Illumina HiSeq2500 platform at the University of Wisconsin-Madison Biotechnology and gene expression center.