title: NextSeq 500 sequencing; Laser Capture Microdissection (LCM) followed by RNA-seq of apical and basal domains of follicle cells from stage 9-10B Drosophila egg chambers
design: Laser Capture Microdissection (LCM) followed by RNA-seq of apical and basal domains of follicle cells from stage 9-10B Drosophila egg chambers
library_construction_protocol: Young (<24h old) w1118 Drosophila females were cultured with standard food supplemented with yeast for 24h before ovary dissection. Usually 10 ovaries (from 10 different flies) were dissected, these were then embedded in cryoblocks and cryosectioned for LCM. RNA from LCM fragments was extracted according to the Geo-seq protocol (Chen et al., Nat Protoc 2017) Sequencing libraries were prepared following the Smart-seq2 protocol described by Picelli et al. (Nat Protoc 2014).