GSM3098213: RNAi_4: OGT RNAi (7 days high sugar diet), replicate 4; Drosophila melanogaster; RNA-Seq
For Gr5a-GAL4/UAS-OGT-RNAi experiments, 20 proboscis per sample were dissected into Trizol (Ambion), and homogenized with plastic pestles. RNA was extracted by acid phenol chloroform (Ambion), and precipitated by isopropanol with Glycoblue Blue Coprecipitant (Invitrogen). RNA pellet was washed as needed with 75% ethanol. RNA was eluted in RNAse DNase free water and treated by DNAse I, following manufacturer's instructions (Turbo DNA-free DNA removal kit, Ambion). For the SD7 and CD libraries, 200 proboscis were dissected in 1xPBS and homogenized in Trizol (Ambion). RNA was extracted by chloroform followed by RNA clean up using RNeasy MinElute Clean Up Kit (Qiagen), and on column DNA digestion by DNAse I (Qiagen). The concentration and integrity of RNA was validated using Agilent Bio-analyzer systems and Qubit RNA High Sensitivity Assay (Invitrogen). Libraries were generated using the Ovation RNA-Seq System for Model Organisms (Nugen, 0350-32) for CD vs SD experiments, and Ovation SoLo RNA-Seq System for Drosophila (Nugen, 0502-96) for Gr5a-GAL4/UAS-OGT-RNAi SD experiments. All reactions included integrated HL-dsDNase treatment (ArcticZymes, Cat. #70800-201).
GEO Accession
GSM3098213