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Sample GSM3475296 Query DataSets for GSM3475296
Status Public on Sep 01, 2019
Title WES_Kit_E211
Sample type SRA
 
Source name Mouse bone marrow cells
Organism Mus musculus
Characteristics strain/background: C57BL/6
genotype/variation: E2-KO
tissue: Bone marrow
cell type: c-Kit+ leukemia cells
Treatment protocol NA
Growth protocol Bone marrow cells were obtained by flushing femurs and tibias with a 25G needle or by crushing femurs, tibias, vertebrae and pelvic bones with a mortar in Ca2+ and Mg2+-free Hank’s buffered salt solution (HBSS; Gibco) supplemented with 2% heat-inactivated bovine serum (HIBS, Gibco). For flow cytometric analysis and isolation, cells were incubated with combinations of antibodies listed in the key resource table. Lineage markers for HSCs and progenitors were CD3, CD5, CD8, B220, Gr1 and Ter119. Antibody staining was performed at 4 °C for 30 min, or, when CD34 was included in the cocktail, on ice for 90 min. Biotinylated antibodies were visualized by incubation with PE/Cy7-conjugated streptavidin at 4°C for 30 min. DAPI (4′,6-diamidino-2-phenylindole; 2 μg/ml in PBS) or PI (propidium iodide; 1 μg/ml) were used to exclude dead cells during flow cytometry.
Extracted molecule genomic DNA
Extraction protocol Genome DNA were isolated from c-Kit+ leukemia cells from E2-KO moribund mice and processed for whole exome sequencing (WES) analysis using Agilent SureSelect XT Mouse All Exon Kit (cat# 1000001746) following manufacturer’s protocols.
 
Library strategy OTHER
Library source genomic
Library selection other
Instrument model BGISEQ-500
 
Data processing Library strategy: WES
Raw reads were mapped to mouse genome mm10 (GRCm38) using SpeedSeq version 0.1.0 with default parameters.
The variants were detected using SpeedSeq version 0.1.0 and VarScan version 2.3.9.
Genome_build: mm10 (GRCm38)
Supplementary_files_format_and_content: VCF format
Supplementary_files_format_and_content: Tab-delimited SNP and Indel calls.
 
Submission date Nov 15, 2018
Last update date Sep 02, 2019
Contact name Jian Xu
E-mail(s) [email protected]
Phone 9015955208
Organization name St. Jude Children's Research Hospital
Department Pathology
Street address 262 Danny Thomas Place, MS 345
City Memphis
State/province Tennessee
ZIP/Postal code 38105
Country USA
 
Platform ID GPL23479
Series (1)
GSE122585 Whole exome sequencing analysis of bone marrow c-Kit+ leukemia cells in Nras-G12D with Ezh2 KO (E2-KO) mice
Relations
BioSample SAMN10431974
SRA SRX5012076

Supplementary file Size Download File type/resource
GSM3475296_WES_Kit_E211_SpeedSeq.vcf.txt.gz 8.7 Mb (ftp)(http) TXT
GSM3475296_WES_Kit_E211_VarScan2.indel.txt.gz 183.8 Kb (ftp)(http) TXT
GSM3475296_WES_Kit_E211_VarScan2.snp.txt.gz 1.1 Mb (ftp)(http) TXT
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file

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