|
| Status |
Public on May 27, 2013 |
| Title |
proB-cell line_Runx1-ERt2_OHT |
| Sample type |
RNA |
| |
|
| Source name |
cell line BMiFLT3(15-3)
|
| Organism |
Mus musculus |
| Characteristics |
cell line: BMiFLT3(15-3)
cell type: proB cells from a murine BCP-ALL
genetic modification: cell transduced with Runx1-ERt2
treatment: OHT
|
| Treatment protocol |
24 hours before sorting the cells were either treated with 0.2 µM OHT to ensure Runx1-ERt2 translocation into the nucleus or treated with the appropriate amount of ethanol as a control. For FACS-sort cells were stained with an antibody against B220. B220 positive cells were isolated and cell pellets were frozen at -80 °C until further processing steps were performed.
|
| Growth protocol |
Cells were routinely grown in α-MEM with 10% FCS, glutamine and sodium pyruvate.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was extracted (NucleoSpin® RNA II; Agilent Technologies), according to the manufacturer's instructions by Miltenyi Biotec.
|
| Label |
Cy3
|
| Label protocol |
Cy3-labeled cRNA was prepared using the Agilent Low Input Quick Amp Labeling Kit (Agilent Technologies) following manufacturer´s protocol. The amount of cRNA and the dye-incorporation rate were checked by using the ND-1000 Spectrophotometer (NanoDrop Technologies) by Miltenyi Biotec.
|
| |
|
| Hybridization protocol |
Hybridization was peformed according to manufacturer´s protocol using the Agilent Gene Expression Hybridization Kit (Agilent Technologies). 1.65 µg Cy3-labeled fragmented cRNA was hybridized overnight (17 hrs., 65 °C) to Agilent Whole Mouse Genome Oligo Microarrays 4x44K by Miltenyi Biotec.
|
| Scan protocol |
Fluorescence signals were scanned using the Agilent´s Microarray Scanner System (Agilent Technologies). The Agilent Feature Extraction Software (EFS) was used to read out and process the microarray image files.
|
| Data processing |
The images were analyzed using the Agilent G2567AA Feature Extraction Software v.9.1.
|
| |
|
| Submission date |
Mar 22, 2013 |
| Last update date |
May 28, 2013 |
| Contact name |
Neele Margarete Kriebitzsch |
| E-mail(s) |
[email protected]
|
| Organization name |
Heinrich-Pette-Institute, Leibniz Institute for Experimental Virology
|
| Lab |
Molecular Pathology
|
| Street address |
Martinistraße 52
|
| City |
Hamburg |
| ZIP/Postal code |
20251 |
| Country |
Germany |
| |
|
| Platform ID |
GPL11202 |
| Series (2) |
| GSE45424 |
Gene expression analysis to identify Runx1 target genes in B-cell progenitors |
| GSE45425 |
Runx1 targets in early B-cell progenitors |
|
