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Status |
Public on Nov 01, 2021 |
Title |
control group |
Sample type |
SRA |
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Source name |
fracture tissues
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Organism |
Mus musculus |
Characteristics |
strain: C57BL/6 tissue: bone fracture tissues( bone and bone marrow) time point: 0 day
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Treatment protocol |
By constructing mouse fracture model, and extracting bone fracture end tissue at different time ( 0day, 3days, 7days, 14days),and extracting single cells by digestion
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Extracted molecule |
total RNA |
Extraction protocol |
To obtain bone and bone marrow cells for scRNA-seq, mice were sacrificed via CO2 asphyxia. Bone fracture tissues (including bones and bone marrow 0.5cm from the fracture end) were dissected from mice and placed in DMEM/F12(Gibco), and cut bone fracture tissues into small fragments and digested them with 0.1mg/ml collagenase II (Invitrogen) for 2h at 37℃. After digestion, the obtained cells were filtered through a 70μm filter and collect it into a collection tube, and erythrocytes lysed in ACK-lysis buffer. RNA libraries were generated by BD Rhapsody singel cell RNA-seq and prepared for sequencing using standard Illumina protocols
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Library strategy |
RNA-Seq |
Library source |
transcriptomic |
Library selection |
cDNA |
Instrument model |
HiSeq X Ten |
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Description |
youer
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Data processing |
Basecalls performed using CASAVA version 1.8 Umi-tools was utilized to calculate the raw counts and cell whitelist by default parameter We applied the Seurat [https://satijalab.org/seurat/] package for cell normalization and cell filtering considering the MT percentage, minimum and maximum gene numbers under following criteria: MT% < 20%; Cell Gene Number < Median Gene Number * 2; Cell Gene Number > 200. PCA and tSNE analysis was used for the single cell to cell relation description. Graphcluster and K-mean was utilized for cell clustering and based on the marker gene achieved some cluster was combined. Wilcox rank sum test was then used for marker gene analysis. Genome_build: mm10 Supplementary_files_format_and_content: TXT file contain counts for each sample
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Submission date |
Jun 18, 2019 |
Last update date |
Nov 01, 2021 |
Contact name |
Renkai Wang |
E-mail(s) |
jfsdfgjh@126.com
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Organization name |
changhai hospital
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Street address |
168 Changhai Road,
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City |
shanghai |
ZIP/Postal code |
200433 |
Country |
China |
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Platform ID |
GPL21273 |
Series (1) |
GSE132884 |
Single-cell RNA-seq analysis reveals two specific endothelial cells promote fracture healing |
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Relations |
BioSample |
SAMN12084343 |
SRA |
SRX6085362 |
Supplementary data files not provided |
SRA Run Selector |
Raw data are available in SRA |
Processed data are available on Series record |
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