|
| Status |
Public on Dec 31, 2012 |
| Title |
Boo Control replicate 4 |
| Sample type |
RNA |
| |
|
| Source name |
Primary culture of c-Kit enriched bone marrow
|
| Organism |
Mus musculus |
| Characteristics |
genotype/variation: Booreana
retrovirus transduction: Control
tissue: Bone marrow
|
| Growth protocol |
Bone marrow cells were isolated from 4 8-week old Booreana and age matched C57Bl6 mice and enriched for c-Kit expressing cells using MACS CD117 microbeads (Miltenyi). Cells were transduced with retroviral supernatants prepared from MSCV-I-GFP and MSCV-AML1-ETO9a-I-GFP. Cells were transduced by retroviral supernatants by spinoculation using fibronectin (Sigma, F-1141) or Retronectin (Takara) using manufacturer’s guidelines. Cells were cultured in IMDM with 20% fetal calf serum and a cocktail of IL-3, IL-6 and SCF cytokines produced in-house. 48 hours post-transduction, cells were sorted for GFP-positive, c-Kit positive cells on a MoFlo (Beckman Coulter) using an anti-c-Kit PECy7 antibody (eBioscience).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was extracted with QIAGEN RNeasy kit in accordance with the prescribed protocol provided with the kit. Quality control was performed with Agilent Bioanalyser.
|
| Label |
biotin
|
| Label protocol |
Illumina TotalPrep RNA Amplification Kit (Ambion) was used to make cRNA
|
| |
|
| Hybridization protocol |
Standard Illumina hybridization protocol
|
| Scan protocol |
Standard Illumina scanning protocol
|
| Description |
BooV4
replicate 4
|
| Data processing |
Variance stabilisation and quantile normalisation using lumi package (R/Bioconductor)
|
| |
|
| Submission date |
Dec 07, 2011 |
| Last update date |
Dec 31, 2012 |
| Contact name |
Konstantin Shakhbazov |
| Organization name |
UQDI
|
| Street address |
Ipswich Road
|
| City |
Brisbane |
| ZIP/Postal code |
4102 |
| Country |
Australia |
| |
|
| Platform ID |
GPL6885 |
| Series (1) |
| GSE34224 |
Interaction of c-Myb with p300 is required for the induction of acute myeloid leukemia (AML) by human AML oncogenes |
|
