title: Spodoptera littoralis adult, Malpighian tubules
design: RNA-seq analysis of S. Litoralis larvae was performed on four separate libraries prepared from RNA isolated from 1) brains, 2) fat bodies, 3) midguts and 4) Malpighian tubules of the last (the sixth) instar larvae. The collections contained pooled tissues from each of five days of the last instar larval development, including freshly molted larvae until prepupal stage. Larvae were kept on artificial diet and fed ad libitum. Colony of insects was reared in the photoperiod of 16h light and 8h darkness and temperature 26oC. Particular tissue samples for each of the four libraries were collected two times of a day, in the middle if the day and in the middle of the night and pooled together for RNA isolation. Approximately 60 tissues from 60 larvae were collected for each library preparation.   The RNA seq analysis on adult moth’s tissues was performed on RNA derived from tissues collected from adult moths Spodoptera littoralis of both sexes, on the second day after eclosion. Four separate libraries were prepared: 1) brains together with optic lobes, 2) fat bodies, 3) midguts and 4) Malpighian tubules.  A colony of insects was reared in the photoperiod of 16h light and 8h darkness and temperature 26oC and butterflies were fed with 10% sucrose solution. Particular tissue samples for each of the four libraries were collected every four hours during the day and pooled together for RNA isolation. Approximately 60 tissues from 60 butterflies were collected for each library preparation.   Material for analysis was snap frozen in liquid nitrogen and stored at -80°C until further use. Total RNA was isolated with standard TRIzol protocol according to manufacturer’s instruction, and RNA integrity was evaluated using a Bioanalyzer 2100 (Agilent). RNA sequencing libraries were prepared with TruSeq Stranded Total RNA Library Prep Kit from Illumina. The 100 bp paired-end Illumina sequencing was conducted at EMBL Genomics Core Facilities (Heidelberg, Germany).