GSM8187272: D56MaleF; Drosophila melanogaster; RNA-Seq
The excretory tract (Malpighian tubules, hindgut and rectal ampulla) was dissected from ice-cold-anaesthetised flies by pulling their intestine from the rectal pad on an RNA stabilisation solution (RNAlater, xxx). The intestine was cut above the insertion of the Malpighian tubules, and the dissected tissue was transferred into 1.5 mL tubes on dry ice (15 samples per tube). RNA quality and concentration was assessed using the Agilent 2100 Bioanalyser RNA 6000 Nano assay. Starting with 200ng Total RNA, polyA enrichment was performed using NEBNext Poly(A) mRNA Magnetic Isolation module, and NGS RNAseq libraries generated using the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina according to manufacturer's instructions. Library quality was assessed using the Agilent 2100 Bioanalyser High-Sensitivity DNA assay For each sample, a minimum of 20 million Paired End 60bp reads (dual 8bp indexing) were generated on an Illumina NextSeq 2000