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Status |
Public on Oct 17, 2017 |
Title |
971672_MB4671F1_S18 |
Sample type |
SRA |
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Source name |
WDR11 Overexpression G3 MB tumorspheres # 19568
|
Organism |
Mus musculus |
Characteristics |
cell description: Mouse MB cells treatment: WDR11 overexpression
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Treatment protocol |
Tumorspheres were dissociated and infected six times over a 2-day period in virus collection media supplemented with N2, B27, EGF and FGF containing lentiviral particles encoding empty vector (pCDH-EF1-MCS-T2A-copGFP) or WDR11 (pCDH-EF1-WDR11-T2A-copGFP. The following day, infected tumorspheres were suspended in Matrigel (BD Bioscience, San Jose) at a concentration of 100,000 tumorspheres/5µl Matrigel and transplanted into the cortices of CD1-nu/nu mice, as described previously [19, 49]. When mice became moribund, tumors were isolated and tumor chunks flash frozen and stored at -80 degrees Celsius.
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Growth protocol |
G3 MB tumorspheres # 19568 were derived from tumors that developed in CD1nu/nu mice implanted with cerebellar granule neuronal progenitors transduced with MSCV retroviruses expressing Myc and the green fluorescent protein (GFP) from an internal ribosomal entry site (IRES) (Myc-IRES-GFP) as previously described (Kawauchi et al, 2012). Briefly, spheres were grown in grown in N2, B27, EGF and FGF supplemented neurobasal medium (“complete tumorsphere medium”).
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Extracted molecule |
total RNA |
Extraction protocol |
Total RNAs were extracted from tumor chunks using Trizol according to manufacturer’s instruction, and as previously published (Murphy et al., 2013). Libraries were prepared from ~500 ng total RNA with the TruSeq RNA Library Preparation Kit v2 according to manufacturer’s instructions (Illumina, San Diego CA)
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Library strategy |
RNA-Seq |
Library source |
transcriptomic |
Library selection |
cDNA |
Instrument model |
Illumina HiSeq 2000 |
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|
Description |
SJMMMB047324
|
Data processing |
Basecalling was performed using Illumina Casava 1.7 FASTQ sequences adapter trimmed (curtadapt) and mapped to the mouse mm9 genome by a pipeline that serially employs STAR and BWA Mapped reads were counted with HTSEQ Genome_build: mm9 Supplementary_files_format_and_content: count files are txt format files that contain raw read counts that map to transcripts for each defined gene.
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Submission date |
Jul 21, 2017 |
Last update date |
May 15, 2019 |
Contact name |
Lei Wei |
E-mail(s) |
lei.wei@roswellpark.rog
|
Phone |
7168451176
|
Organization name |
Roswell Park Cancer Institute
|
Department |
Biostatistics & Bioinformatics
|
Street address |
Elm & Carlton Streets
|
City |
Buffalo |
State/province |
NY |
ZIP/Postal code |
14263 |
Country |
USA |
|
|
Platform ID |
GPL13112 |
Series (1) |
GSE101767 |
Exome sequencing analysis of murine medulloblastoma models identifies Wdr11 as a potential tumor suppressor in Group 3 tumors |
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Relations |
BioSample |
SAMN07376336 |
SRA |
SRX3028088 |