title: RNA-seq of adult midguts of wts RNAi-2
design: To extract total RNAs for RNA-Seq experiment, we used 30 midguts dissected from Actin-Gal4; tub-Gal80ts (Actints, control), Actints > wts RNAi (wts RNAi), Actints > lola RNAi (lola RNAi), Actints > yki (yki) flies incubated for 5 days at 29C per sample. Invitrogen TRIzol Reagent (15596026, Invitrogen) is used to isolate total RNA according to the User Guide. After assessing RNA quality with Agilent Bioanalyzer, mRNAs were enriched by poly-A pull-down from total RNA samples. Then, sequencing libraries constructed with Illumina TruSeq Stranded mRNA Sample Prep Kit were sequenced using Illumina HiSeq X ten.