RNA-Seq of Trichomalopsis sarcophagae male pupa
T. sarcophagae were collected at five life-stages: 0-10hr early embryo, 14-24hr late embryo, 44-54hr larva, yellow pupa, and adults. Six individuals were pooled per stage, except early embryo for which ~40 individuals were pooled due to small size. Samples were collected on dry ice, homogenized in 100ul TRIzol with an additional 900ul TRIzol was added, and stored at -80oC. Total RNA extractions were performed as per manufacturerŐs protocol, followed by quantification and quality checking using Agilent 2100 Bioanalyzer. TruSeq mRNA (Illumina) library construction and 100bp single-end sequencing on Illumina HiSeq 2500 platform were performed by University of Rochester Genomics Research Center (URGRC). cDNA for each species was indexed with a unique adapter. Each library was normalized by equimolar multiplexing before sequencing at ~ 1 library/10th of a lane.