GSM6788526: brat-KD, rep3; Drosophila melanogaster; RNA-Seq
brains were manually dissected from 96h-old larvae & dissociated into individual cells; total RNA from GFP-positive cells (type 2 neuroblasts & their descendents) was sorted by FACS, RNA was isolated using Agilent Technologies' Absolutely RNA Nanoprep Kit (including DNase I digestion) and RNA concentration and quality were determined on a 2100 Bioanalyzer Instrument (Agilent Technologies) using the Agilent RNA 6000 Pico Kit (Agilent Technologies) Library preparation was performed using the Poly(A) mRNA Magnetic Isolation Module (New England Biolabs) and the NEBNext Ultra II Directional RNA library Prep Kit for Illumina (New England Biolabs). For library amplification 17 or 24 PCR cycles were used. Library size distribution and concentration were analyzed on the Fragment Analyzer (Agilent Technologies) using the NGS Fragment High Sensitivity Analysis Kit (1-6,000 bp; Agilent Technologies). The libraries were sequenced on the NextSeq500 Illumina platform for 75 cycles. Base calling was performed using Illumina's BaseSpace platform