|
| Status |
Public on Sep 01, 2018 |
| Title |
MIE rep2 |
| Sample type |
RNA |
| |
|
| Source name |
Empty EGFP vector (MIE) transduction of murine hematopoietic stem cells
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL/6
tissue: Blood
cell type: HSC
genotype: control
|
| Treatment protocol |
The bone marrow cells were infected with MIE, WT and R396Q retroviral particles. One hundred thousand cells per condition were infected twice with retroviral vector-based particles by centrifugation at 3,000xg for 2 hours at 32°C with 2µg Polybrene (Hexadimethrine bromide, 400µg/uL) on 2 consecutive days. GFP-positive cells were sorted after 72h of culture.
|
| Growth protocol |
C57BL/6 8-week female were treated with 5-fluorouracil (5-FU) at 150mg/kg 6 days prior to bone marrow cell harvesting. Bone marrow cells from C57BL/6 8-week female mice were sorted according to stem cell surface markers (Lin-Sca+Kit+) using autoMACS (Miltenyi Biotec, Bergisch Gladbach, Germany).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using Rneasy plus MiniKit (Qiagen) and its quality was assessed with the RNA Screen Tape on Agilent 2200 Tapestation Bioanalyzer (Agilent Technologies).
|
| Label |
Biotin
|
| Label protocol |
RNA samples were purified and prepared according to the manufacturer's protocol using the Affymetrix's GeneChip Whole Transcript Sense Target Labeling Assay Kit (Affymetrix) and hybridized on Affymetrix GeneChip Mouse Exon 1.0 ST arrays.
|
| |
|
| Hybridization protocol |
standard Affymetrix protocol
|
| Scan protocol |
standard Affymetrix protocol
|
| Data processing |
Probe-signal intensities were normalized and summarized using the robust multiarray average (RMA) method with the Affymetrix’s Expression Console software.
affymetrix-algorithm-name = sst-rma-gene-full
affymetrix-algorithm-version = 1.0
|
| |
|
| Submission date |
Mar 02, 2018 |
| Last update date |
Sep 01, 2018 |
| Contact name |
Eric DELABESSE |
| E-mail(s) |
delabesse.e@chu-toulouse.fr
|
| Organization name |
Institut Universitaire de Cancérologie de Toulouse, site Oncopôle CHU de Toulouse
|
| Street address |
1 avenue Irène Joliot-Curie
|
| City |
Toulouse |
| ZIP/Postal code |
31059 |
| Country |
France |
| |
|
| Platform ID |
GPL20258 |
| Series (1) |
| GSE111328 |
GATA2 mutant transduction of murine hematopoietic stem cells |
|
