GSE136676
The RNA-seq analyses of transcriptional changes responsing to manganese depletion in Drosophila
Purpose: identifying genes responding to manganese depletion in Drosophila through whole transcriptome RNA-seq analyses Methods: 10 females and 10 males of wild type of the Oregon R-C genotype, were mated and allowed to lay eggs for five days on two variants of chemical diet (no manganese added or 5 ppm manganese). Parents were removed and recently-eclosed females were transferred to fresh medium, respectively, for another five days. 10 midguts were dissected in PBS and homogenized in TRIzol (Invitrogen). RNA extraction and purification was performed using the Direct-zol RNAMicroPrepkit (Zymo Research) method.  two biological replicates were used for each dietary treatment group. The RNA was eluted with sterile water and each RNA solution was divided into two aliquots, one of which was freshly used to obtain an RNA Integrity Number (RIN) evaluation at the Harvard Medical School Biopolymers Facility. For samples with RIN > 6.7 and no RNA degradation, the other aliquot stored at -80°C was submitted for sequencing. Results: we identified differentially expressed genes responding to to manganese depletion in Drosophila. Overall design: Samples of two chemical diets (no manganese or 5 ppm manganese added) with replicates were sequenced and analyzed for differentially expressed genes responsing to manganese depletion.