GSM6799486: EB Ctrl rep 3; Drosophila melanogaster; RNA-Seq
RNA were purified from each cell sample using MasterPure RNA purification kit (Epicentre), following manufacturer's recommendation. Sorted cells were incubated at 65°C with rocking for 15 minutes. Samples were put on ice then total nucleic acid were precipitated. Contaminating DNA was removed using DNase, then nucleic acids were precipitated again. RNAs were finally resuspended in 10uL TE and 1uL RiboGuard (RNases inhibitor) and stored at -20°C. Construction of RNA banks and sequencing was made at the Montpellier GenomiX Platform (MGX). RNA banks were obtained using the Ovation SoLo RNAseq System kit (Nugen). Bank validation was done with the quantification of the complementary DNA with the Standard Sensitivity NGS kit on Fragment Analyzer and with qPCR (ROCHE Light Cycler 480). 100nt single-reads sequencing was performed on NovaSeq 6000 (Illumina) with NovaSeq Reagent Kits (100cycles).