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| Status |
Public on Aug 13, 2018 |
| Title |
Fev-Cre;mTmG reporter |
| Sample type |
SRA |
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| Source name |
pancreatic cells
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| Organism |
Mus musculus |
| Characteristics |
strain: Fev-Cre;ROSA26mTmG
tissue: pancreas
developmental stage: E14.5
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| Extracted molecule |
total RNA |
| Extraction protocol |
Pancreata were isolated from mouse embryos and dissociated in TrypLE Express at 37C with pipet trituration at 5-minute intervals during incubation. E12.5 pancreata were dissociated for 10 minutes, E14.5 pancreata for 15 minutes, and E17.5 pancreata for 30 minutes. Dissociations were neutralized with FACS buffer (10% FBS + 2mM EDTA in phenol-red free HBSS). Dissociated cells were passed through a 30um cell strainer and stained with Sytox live/dead stain (Thermo Fisher), along with protocol specific stains noted in the individual sample extract protocol descriptions. Stained cells were washed twice in FACS buffer and then processed through a BD FACS Aria II.
For E14.5 Fev-Cre;ROSA26mTmGpancreata, live GFP+ and Tdtomato+ cells were collected in separate tubes.
cDNA libraries were prepared for sequencing using standard protocol for the 10x Single Cell instrument with v2 kit and reagents.
droplet-based single cell RNA-Seq
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| Library strategy |
RNA-Seq |
| Library source |
transcriptomic |
| Library selection |
cDNA |
| Instrument model |
Illumina NovaSeq 6000 |
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| Description |
single cell RNA sequencing of E14.5 Fev-Cre;mTmG pancreas, aligned to mm10 genome with GFP and TdTomato added
Read 1= 98, Index read 1 (i7)= 14, Index read 2 (i5)= 8, Read 2= 10
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| Data processing |
Data were processed according to the 10x Cellranger pipeline.
BAM files were generated with v2.1 10X Genomics Cellranger software using default parameters.
Gene-barcode matrices were generated from Cellranger v2.1 using default parameteres.
Genome_build: mm10 for all Swiss Webster. For Fev-Cre;Rosa26mTmG, eGFP and TdTomato sequences from the mTmG mouse line (Muzumdar MD. et. al., 2007) were added to the mm10 genome FASTA and annotations. The genome was prepared as according to 10X Genomics instructions for custom genome builds.
Supplementary_files_format_and_content: Cellranger outputs a .mtx file with the gene-barcode matrix. This matrix contains the counts of molecules per cell (UMI/cell) as determined after filtering and counting by Cellranger. The matrix is in market exchange format while the row (gene names) and columns (cell barcodes) are provided as TSV files. File formats are documented in Cellranger software manual.
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| Submission date |
May 14, 2018 |
| Last update date |
Aug 14, 2018 |
| Contact name |
Jeremy Reiter |
| E-mail(s) |
[email protected]
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| Organization name |
UCSF
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| Street address |
555 Mission Bay Boulevard South
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| City |
San Francisco |
| State/province |
California |
| ZIP/Postal code |
94158 |
| Country |
USA |
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| Platform ID |
GPL24247 |
| Series (1) |
| GSE101099 |
Lineage dynamics of pancreatic development at single cell resolution |
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| Relations |
| BioSample |
SAMN09205833 |
| SRA |
SRX4080250 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSM3140920_Fev_Cre_barcodes_reporter.tsv.gz |
30.9 Kb |
(ftp)(http) |
TSV |
| GSM3140920_Fev_Cre_genes_reporter.tsv.gz |
217.5 Kb |
(ftp)(http) |
TSV |
| GSM3140920_Fev_Cre_reporter.mtx.gz |
69.6 Mb |
(ftp)(http) |
MTX |
SRA Run Selector |
| Processed data provided as supplementary file |
| Raw data are available in SRA |
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