|
| Status |
Public on Oct 06, 2006 |
| Title |
BM CD34+ Pool I |
| Sample type |
RNA |
| |
|
| Source name |
Bone Marrow CD34+ progenitors cells
|
| Organism |
Homo sapiens |
| Characteristics |
CD34+ hemopoietic progenitors purified from bone marrow of healthy controls
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using Trizol (Invitrogen-Life Technologies, Paisley, UK). Disposable RNA chips (Agilent RNA 6000 Nano LabChip kit, Agilent Technologies, Waldbrunn, Germany) were used to determine the concentration and purity/integrity of RNA with the Agilent 2100 Bioanalyzer. In order to obtain enough RNA to perform the Affymetrix analysis, 0.6 µg of RNA from CD34+ cells of 5 different normal donors or IM patients were pooled, so as to obtain 3 pools each comprising 3 µg RNA.
|
| Label |
biotin-labeled
|
| Label protocol |
Biotin-labeled labeled cRNA was purified using RNeasy spin columns (Qiagen, Valencia, CA) and fragmented (20 µg) following the Affymetrix GeneChip protocol. The assessment of cRNA concentration/quality and fragmentation was performed with the Agilent RNA chips.
|
| |
|
| Hybridization protocol |
The fragmented cRNA was then hybridized to an identical lot of Affymetrix HG-U133A GeneChip arrays for 16 hours. GeneChips were washed and stained using the instrument’s standard Eukaryotic GE WS2v4 protocol, using antibody-mediated signal amplification.
|
| Scan protocol |
GeneChip were finally scanned using the GA2500 Affymetrix GeneChip scanner.
|
| Description |
The biotin-labeled target synthesis reactions, as well as the Affymetrix HG-U133A GeneChip arrays hybridization, staining, and scanning, were performed using Affymetrix standard protocols (Affymetrix, Santa Clara, CA).
|
| Data processing |
GeneChip Operating Software (GCOS)
|
| |
|
| Submission date |
Oct 04, 2005 |
| Last update date |
Oct 06, 2006 |
| Contact name |
Rossella Manfredini |
| E-mail(s) |
manfredini.rossella@unimore.it
|
| Phone |
+390592058065
|
| Organization name |
Centre for Regenerative Medicine
|
| Department |
Life Sciences
|
| Street address |
Via Gottardi 100
|
| City |
Modena |
| ZIP/Postal code |
41100 |
| Country |
Italy |
| |
|
| Platform ID |
GPL96 |
| Series (1) |
|
