RNA-Seq of Drosophila: whole body strain w1118;TBPH23/23
cDNA libraries were constructed from450ngof total RNA by using theQuantSeq 3 mRNA-Seq Library Prep Kit for Illumina (FWD)(Lexogen, Vienna, Austria) according to the manufacturers instructions. The workflow consists offirst strand cDNA synthesis with oligo(dT) primers containing an Illumina-compatible sequence at the 5 end, RNA template removal, second strand synthesis with random primers containing an Illumina-compatible linker sequence at the 5 end, purification using magnetic beads to remove all reaction components, and PCR amplification in order to add the complete adapter sequences and to generate the final library. The libraries were quantified with theQubitFlexFluorometer(Invitrogen, Carlsbad, USA) and quality tested byAgilent 4150 TapeStation system(Agilent Technologies, Santa Clara, USA). Sequencing was carried out insingle-endmode (150 bp) by usingNovaSeqX Plus(Illumina, San Diego, USA) with a targeted sequencing depth of30 millionreads per sample. Base-calling was performed using RTA2 software (Illumina, San Diego, California, USA).