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SRX7814226: GSM4363298: WT Replicate 1; Drosophila melanogaster; RNA-Seq
8 ILLUMINA (Illumina NovaSeq 6000) runs: 429.9M spots, 50.3G bases, 14.6Gb downloads

Submitted by: NCBI (GEO)
Study: Single-cell RNA sequencing of adult Drosophila ovary identifies transcriptional programs governing oogenesis
show Abstracthide Abstract
Using single-cell RNA sequencing technology, we have built a comprehensive transcriptomic dataset for the adult Drosophila ovary and connected tissues. A total of 429,855,892 reads were obtained with 28,995 mean reads per cell for the final dataset of 7,053 high-quality cells containing an overall 11,782 genes. Using this dataset, we identified the transcriptional trajectory of the entire follicle cell population over the course of their development from stem cells to the oogenesis-to-ovulation transitioning phase in the Corpus luteum. We further identify expression patterns during essential developmental events which take place in somatic and germline cell types such as differentiation, cell-cycle switching, migration, symmetry breaking, nurse cell engulfment, egg-shell formation, and corpus luteum signaling. Extensive experimental validation of unique expression patterns in both ovarian and non-ovarian cells also led to the identification of many new markers specific to cell-type and stage. The inclusion of several nearby tissue types in this dataset also led to our identification of functional convergence in expression between distantly related cell types such as the immune-related genes which were similarly expressed in immune cells (hemocytes) and ovarian somatic cells (stretched cells) during their brief phagocytic role in nurse cell engulfment. Taken together, these findings provide new insight into the temporal regulation of genes in a cell-type specific manner during oogenesis and begin to reveal the relatedness in expression between cell and tissues types. Overall design: scRNA-seq (10x Genomics CHROMIUM Single Cell 3' Solution V2 Chemistry and Illumina NovaSeq 6000) of adult ovaries from female Drosophila melanogaster flies.
Sample: WT Replicate 1
SAMN14237454 • SRS6229459 • All experiments • All runs
Library:
Instrument: Illumina NovaSeq 6000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: Whole ovaries were extracted from female flies that were dissected in complete medium (Grace's Insect Basal Medium supplemented with 15% FBS) and were transferred to a tube containing 300 μL EBSS (no calcium, magnesium, and phenol red), followed by a gentle wash for 2 minutes. The EBSS was then removed and the tissue was dissociated in 100 μL Papain (50 U/mL in EBSS and previously heat activated in 37°C for 15 minutes) for 30 minutes. The suspension was mechanically dissociated every 3 minutes by gentle pipetting up and down. To quench the digestion, 500 μL complete medium was added to dissociated cells. The suspension was then passed through a 40 μL sterile cell strainer and centrifuged for 10 minutes at 700 RCF to remove large eggs with intact eggshell which cannot be dissociated and debris. Supernatant was removed and single cells were re-suspended in 100 μL. Cell viability was assayed using Trypan Blue and estimates of cell concentration were made using a hemocytometer. Cells were then further diluted to an approximate, final concentration of 2,000 cells/μL according to 10X Genomics recommendations. Single-cell libraries were prepared using the Single Cell 3' Library & Gel Bead Kit v2 and Chip Kit according to the recommended 10X Genomics protocol. Single cell suspension was loaded onto the Chromium Controller (10X Genomics). Library quantification assays and quality check analysis was performed using the 2100 Bioanalyzer instrument (Agilent Technologies). The library samples were then diluted to a 10nM concentration and loaded onto two lanes of the NovaSeq 6000 (Illumina) instrument flow cell for a 100-cycle sequencing run.
Experiment attributes:
GEO Accession: GSM4363298
Links:
Runs: 8 runs, 429.9M spots, 50.3G bases, 14.6Gb
Run# of Spots# of BasesSizePublished
SRR1121557151,720,4286.1G1.8Gb2020-03-27
SRR1121557252,007,2406.1G1.8Gb2020-03-27
SRR1121557358,517,5116.8G2Gb2020-03-27
SRR1121557458,900,7186.9G2Gb2020-03-27
SRR1121557540,015,9534.7G1.3Gb2020-03-27
SRR1121557640,302,6604.7G1.4Gb2020-03-27
SRR1121557763,995,3237.5G2.2Gb2020-03-27
SRR1121557864,396,0597.5G2.2Gb2020-03-27

ID:
10218837

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