|
| Status |
Public on Jan 13, 2010 |
| Title |
Patient_45_6hr_AS602868_10µM_rep1 |
| Sample type |
RNA |
| |
|
| Source name |
Leukemic blasts, 6hr, AS602868, 10µM, repilicate 1
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: leukemic blast
gender: m
age: 34y
|
| Treatment protocol |
Isolated leukemic blasts from AML patients were incubated in RPMI 1640 medium supplemented with 10% heat-inactivated fetal bovine serum in presence or absence of 10 µM AS602868 for 6 hours at 37 ºC in a humidified incubator with 5% CO2
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was prepared using the RNeasy Mini Kit (Qiagen, Hilden, Germany) according to the manufacturer’s recommendations and quantified on a Nanodrop ND-1000 spectrophotometer (Nanodrop Technologies, Wilmington, DE, USA). RNA integrity was verified using the BioAnalyser 2100™ (Agilent Technologies, Palo Alto, CA, USA).
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 0.5 ug RNA using the One-Color Low RNA Input Linear Amplification PLUS kit (Agilent) according to the manufacturer's instructions, followed by RNAeasy column purification (QIAGEN, Valencia, CA). Dye incorporation and cRNA yield were checked with the NanoDrop ND-1000 Spectrophotometer
|
| |
|
| Hybridization protocol |
Cy3-labelled cRNA was fragmented at 60 °C for 30 minutes in a reaction volume of 55 µl containing 1x Agilent fragmentation buffer and 2x Agilent blocking agent following the manufacturers instructions. On completion of the fragmentation reaction, 55 µl of 2x Agilent hybridization buffer was added to the fragmentation mixture and hybridized to Whole Human Genome 4 x 44K microarrays (G4112F) for 17 hours at 65 °C in a rotating Agilent hybridization oven. After hybridization, microarrays were washed 1 minute at room temperature with GE Wash Buffer 1 (Agilent) and 1 minute with 37 °C GE Wash buffer 2 (Agilent).
|
| Scan protocol |
Slides were scanned immediately after washing on the Agilent DNA Microarray Scanner (G2505B) with default settings, and data extracted by Feature Extraction software, version 9.5.
|
| Description |
Gene expression after 6h exposure of human leukemic blasts to 10 µM AS602868
|
| Data processing |
Data were normalized using the quantile method
|
| |
|
| Submission date |
Jan 12, 2010 |
| Last update date |
Jan 12, 2010 |
| Contact name |
Lars Petter Jordheim |
| E-mail(s) |
lars.jordheim@recherche.univ-lyon1.fr
|
| Organization name |
INSERM U590
|
| Street address |
8 Av Rockefeller
|
| City |
Lyon |
| ZIP/Postal code |
69008 |
| Country |
France |
| |
|
| Platform ID |
GPL4133 |
| Series (1) |
| GSE19853 |
Gene expression profile of fresh human acute myeloid leukemia cells exposed ex vivo to AS602868 |
|
