|
Status |
Public on Nov 01, 2011 |
Title |
SRBCT biological rep2 |
Sample type |
RNA |
|
|
Source name |
Small Round Blue Cell Tumor
|
Organism |
Danio rerio |
Characteristics |
tissue: tumor genotype/variation: Tg(HSP:EWS-FLI1); p53M214K/M214K
|
Treatment protocol |
Fish were grown under standard conditions and monitored for tumor development. Fish developing tumors were euthanized and the tumor tissue removed for histology and RNA preparation.
|
Growth protocol |
Wild-type or p53(M214K) homozygous zebrafish embryos were injected at the one cell stage with a transposon in which the Heat Shock Protein 70 (HSP70) promoter drove expression of a human EWS-FLI1 fusion oncoprotein [Tg(HSP:EWS-FLI1)].
|
Extracted molecule |
total RNA |
Extraction protocol |
Total RNA was extracted from tumors using Trizol reagent according to the manufacturer's instructions.
|
Label |
biotin
|
Label protocol |
Total RNA (2.5ug) from tumors was reverse-transcribed using a T7-Oligo(dT) Promoter Primer, followed by RNAseH-mediated second-strand cDNA synthesis. Biotinylated cRNA probe was prepared with MessageAmp II (Ambion).
|
|
|
Hybridization protocol |
Hybridization was carried out according to Affymetrix protocol 701027 rev 5, using Affymetrix Fluidics Station 450.
|
Scan protocol |
Arrays were scanned with the Affymetrix GeneChip Scanner 3000 using default parameters
|
Description |
Ewing's Sarcoma-like small round blue cell tumor arising in Tg(HSP:EWS-FLI1); p53M214K/M214K zebrafish
|
Data processing |
The probe-level data were background adjusted, normalized and summarized by a Robust Multi-array Average (RMA) algorithm implemented in the software package affy of the Bioconductor project release 1.7, running in the R statistical environment version 2.2.0 using default parameters.
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|
|
Submission date |
Aug 03, 2011 |
Last update date |
Nov 01, 2011 |
Contact name |
James F Amatruda |
E-mail(s) |
[email protected]
|
Phone |
214-648-3896
|
Organization name |
University of Texas Southwestern Medical Center
|
Department |
Pediatrics and Molecular Biology
|
Street address |
5323 Harry Hines Boulevard
|
City |
Dallas |
State/province |
TX |
ZIP/Postal code |
75390-8534 |
Country |
USA |
|
|
Platform ID |
GPL1319 |
Series (2) |
GSE31186 |
The human Ewing's Sarcoma oncoprotein EWS-FLI1 causes Ewing's-type tumors in zebrafish |
GSE31217 |
A zebrafish transgenic model of Ewing's Sarcoma reveals conserved mediators of EWS-FLI1 tumorigenesis |
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