|
| Status |
Public on Sep 22, 2006 |
| Title |
B-CLL sample 708, good prognosis |
| Sample type |
RNA |
| |
|
| Source name |
immunomagnetically purified fresh CD19+ peripheral blood CLL cells
|
| Organism |
Homo sapiens |
| Characteristics |
immunomagnetically purified native B-CLL cells from the peripheral blood of untreated patients
|
| Treatment protocol |
no treatment
|
| Growth protocol |
no in vitro culture
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Isolation of total RNA was performed with QIAGEN Rneasy columns according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 6 microg total RNA (Expression Analysis Technical Manual, 2001, Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 microg of cRNA were hybridized for 16 hr at 45C on HG-U133A Arrays. GeneChips were washed and stained in the Affymetrix Fluidics Station 400.
|
| Scan protocol |
GeneChips were scanned using the Hewlett-Packard GeneArray Scanner G2500A.
|
| Description |
Aim: to compare the transcriptosomes of good prognosis CLL cases (ZAP-70-CD38-) to poor prognosis cases (ZAP-70+CD38+). Methods: Immunomagnetic isolation was done using MidiMacs resulting in >95% purity of leukemic cells as detected by FACS analysis of CD19+CD5+ cells. The leukemic cells were freshly purified from untreated patients and RNA was directly isolated from fresh cells without further ex vivo treatment of the cells.Eight immunomagnetically purified peripheral blood derived ZAP-70+CD38+ CLL cases were compared with eight ZAP-70-CD38- B-CLL cases.
|
| Data processing |
The data were analyzed with Microarray Suite version 5.0 (MAS 5.0) using Affymetrix default analysis settings and global scaling as normalization method. The trimmed mean target intensity of each array was arbitrarily set to 1000.
|
| |
|
| Submission date |
Mar 07, 2006 |
| Last update date |
Sep 22, 2006 |
| Contact name |
Ludger Klein-Hitpass |
| E-mail(s) |
ludger.klein-hitpass@uni-essen.de
|
| Phone |
+49 201 723 85552
|
| Organization name |
Institut fuer Zellbiologie
|
| Department |
Universitaetsklinikum
|
| Lab |
BioChip Lab
|
| Street address |
Virchowstr. 173
|
| City |
Essen |
| ZIP/Postal code |
D-45122 |
| Country |
Germany |
| |
|
| Platform ID |
GPL96 |
| Series (1) |
| GSE4392 |
Expression data from native ZAP-70+CD38+ vs. ZAP-70-CD38- CLL cells |
|
