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| Status |
Public on Dec 19, 2019 |
| Title |
A cell atlas of the adult Drosophila midgut |
| Organism |
Drosophila melanogaster |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Drosophila guts were dissociated to single cells as previously described (Dutta et al., 2015) with a few modifications. Guts were dissected from 7-day old adult esg-sfGFP/+, pros- Gal4>RFP/+ females. After pulling out the gut, the crop and midgut/hindgut junction (where the Malpighian tubules branch out of the gut) and Malpighian tubules were removed. Five flies at a time were dissected and immediately transferred into cold PBS containing 1% BSA to avoid exposing the midgut tissue to room temperature for a long period of time. Once 40 guts were dissected, they were transferred to a dissection plate and chopped into small pieces using a razor blade. These small fragments were immediately transferred to an eppendorf tube containing 400 ul of 1 mg/ml elastase/PBS solution (Sigma E0258) and incubated on a shaker at 27 ̊C for 30 min. 1% of BSA in PBS (final concentration) was used to stop the digestion reaction and prevent cells from aggregating. The cell suspension was passed through 100 um and then a 40 um cell strainer and loaded on the top of Optiprep with density gradient 1.12 g/ml. Viable cells were isolated from the top layer of the sample after centrifugation at 800xg for 20 min. Cell viability and number were assessed by 0.4% trypan blue and hemocytometer.
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| Overall design |
inDrop and 10x single-cell RNA-sequencing of 10605 cells from the Drosophila midgut
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| Contributor(s) |
Hung R, Hu Y, Kirchner R, Li F, Xu C, Comjean A, Tattikota SG, Song WR, Sui SH, Perrimon N |
| Citation(s) |
31915294 |
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| Submission date |
Sep 26, 2018 |
| Last update date |
Dec 17, 2020 |
| Contact name |
Rory Kirchner |
| E-mail(s) |
[email protected]
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| Organization name |
Harvard T.H. Chan School of Public Health
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| Department |
Department of Biostatistics
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| Street address |
677 Huntington Ave
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| City |
Boston |
| State/province |
MA |
| ZIP/Postal code |
02115 |
| Country |
USA |
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| Platforms (1) |
| GPL19132 |
Illumina NextSeq 500 (Drosophila melanogaster) |
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| Samples (4)
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| Relations |
| BioProject |
PRJNA493298 |
| SRA |
SRP162698 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE120537_counts.csv.gz |
8.1 Mb |
(ftp)(http) |
CSV |
| GSE120537_metadata-code-book.md.txt.gz |
377 b |
(ftp)(http) |
TXT |
| GSE120537_metadata.csv.gz |
746.7 Kb |
(ftp)(http) |
CSV |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
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