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Sample GSM5915204 Query DataSets for GSM5915204
Status Public on Nov 08, 2023
Title Head_RNA_DD25-2
Sample type SRA
 
Source name Male adult heads
Organism Drosophila melanogaster
Characteristics strain: w1118
photoperiod: Consistent darkness
age after eclosion: 25 days
replicate: 2
Treatment protocol We transferred these 1-day-old adults to a climatic chamber under the photoperiodic cycles of BD and DD, respectively. We collected the bulk 10- and 25-day-old male adult heads (BD10, BD25, DD10, and DD25) for poly-A RNA-seq.
Growth protocol We collected the eggs oviposited by 3-day-old Drosophila melanogaster w1118 adults which were colonized under the photoperiodic cycles of 12 h white fluorescent light: 12 h darkness (LD) after eclosion. These eggs were grown on standard mediums (comprised of 80 g cornmeal, 125 g sucrose, 16 g yeast, and 5 g agar/L) under the photoperiodic cycles of LD until one day after eclosion.
Extracted molecule polyA RNA
Extraction protocol Total RNAs were isolated and dissolved in RNase-free water after the bulk samples had been homogenized in 0.5–1 mL TRIzol AG RNAex Pro Reagent (#AG21102, Accurate Bioeng. Co., Ltd., Hunan, China) with glass beads using a homogenizer.
Poly-A RNAs were purified from the extracted total RNAs using Dynabeads™ Oligo(dT)25 (#61002, Invitrogen, CA, USA) and were further fragmented as ~350 nt fragments using NEBNext® Second Strand Synthesis (dNTP-Free) Reaction Buffer (#B6117S, New England BioLabs, MA, USA). Strand-specific sequencing libraries were conducted using NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (#E7530L, New England BioLabs) according to the manufacturer's protocols including first- and second-strand cDNA synthesis, adaptor ligation, and library amplification with index sequences.
 
Library strategy RNA-Seq
Library source transcriptomic
Library selection cDNA
Instrument model Illumina NovaSeq 6000
 
Description Parental photoperiod: White-light: darkness = 12 h: 12h; Parental age after eclosion: 3 days
Data processing After removing adaptors and low-quality bases by fastp (version 0.20.1), the resulting reads no less than 50 bp were mapped to the Drosophila melanogaster reference genome (FB2021_02) released by FlyBase database (http://flybase.org/).
The featureCounts program within SourceForge Subread package (version 2.0.2) was used for counting the reads that mapped to genes.
The genes with an average read count less than 10 in any group or lacking read count in any replicate were primarily filtered.
R package DESeq2 (version 1.30.0) was used for the identification of differentially expressed genes (DEGs) by setting false discovery rate corrected P-value (FDR) < 0.05 and fold change >= 2 as the thresholds of significance (Supplementary Table S2), and the read count normalization across samples by size factors.
t-SNE dimension reduction analysis was performed using R (version 4.0.3) package Rtsne (version 0.15); heatmap was generated using R package pheatmap (version 1.0.12); common and unique DEGs were visualized in a Venn diagram generated using R package VennDiagram; volcano plot was generated using R package ggplot2; GO (Gene Ontology) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses for the DEGs were performed using either Metascape (version 3.5) (https://metascape.org/) or R package clusterProfiler (version 3.18.1); Gene set enrichment analysis (GSEA) software (version 4.2.0) was used for determining the statistical significance of priori defined gene sets.
Genome_build: dm6
Supplementary_files_format_and_content: Tab-delimited text file (.txt) includes raw read counts for the RNA-seq data of w1118 male adult heads.
 
Submission date Feb 24, 2022
Last update date Nov 08, 2023
Contact name Xiaoyun Wang
E-mail(s) wang_xiaoyun@gibh.ac.cn
Organization name Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences
Department Infection and Immunity
Street address 190 Kaiyuan Avenue, Huangpu
City Guangzhou
State/province Guangdong
ZIP/Postal code 510530
Country China
 
Platform ID GPL25244
Series (2)
GSE197370 5' UTR-dependent m6A methylation regulates aging and circadian rhythms in Drosophila [RNA-seq head]
GSE197397 5' UTR-dependent m6A methylation regulates aging and circadian rhythms in Drosophila
Relations
BioSample SAMN26232704
SRA SRX14279276

Supplementary data files not provided
SRA Run SelectorHelp
Raw data are available in SRA
Processed data are available on Series record

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