|
| Status |
Public on Mar 01, 2011 |
| Title |
B-CLL PB P137 |
| Sample type |
RNA |
| |
|
| Source name |
B-CLL PB P137
|
| Organism |
Homo sapiens |
| Characteristics |
disease state: B-CLL
tissue: peripheral blood
|
| Growth protocol |
Peripheral blood from B-CLL patients as well as from healthy donors were firstly separated by Ficoll gradient (GE Healthcare), obtained peripheral mononuclear cells (PBMC). PBMC were shortly incubated in IMDM (Isove’s Modified Dulbecco’s) medium supplemented with 10% fetal calf serum, 1% non essential amino acids, 1% antibiotics. PBMCs from healthy donors were separated by autoMACS Pro Separator and anti FITC microbeads, CD19 human FITC antibody were used.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted by TRIzol Reagent following the manufacturer protocol. Precipitation by isopropyl alcohol (0.5mL per 1mL TRIzol) was done over night. As carrier of the aqueous phase the Acrylamide was used. The RNA pellets were washed with 75% ethanol (1mL per 1mL TRIzol) and dry pellets were resuspend in 20 uL of RNAse free water.
|
| Label |
biotin
|
| Label protocol |
RNA was proceeded according to Affymetrix 3‘ – IVT Express Kit protocol. cDNA were prepared using the recommended Affymetrix reagents and kits.
|
| |
|
| Hybridization protocol |
RNA was proceeded according to Affymetrix 3‘ – IVT Express Kit protocol. cDNA was hybridized for 16 hr to the Affymetrix array as described in the Expression Analysis Technical Manual. GeneChips were washed and stained in the Affymetrix Fluidics Station.
|
| Scan protocol |
GeneChips were processed using the Agilent GeneArray Scanner 3000 7G system in the Genomics Facility of the New York University Cancer Institute.
|
| Description |
Additional information and characteristics in the referred paper supplement xls table.
|
| Data processing |
Data were summarized using GeneSpring implementation of RMA algorithm and expression data were normalized on median of control samples.
|
| |
|
| Submission date |
Jan 19, 2011 |
| Last update date |
Mar 01, 2011 |
| Contact name |
Vojtěch Kulvait |
| E-mail(s) |
vojtech.kulvait@lf1.cuni.cz
|
| Phone |
+420224965968
|
| Organization name |
Charles University in Prague - 1st Faculty of Medicine
|
| Department |
Institute of Pathological Physiology
|
| Lab |
Centrum for experimental haematology
|
| Street address |
U Nemocnice 5
|
| City |
Prague 2 |
| ZIP/Postal code |
128 53 |
| Country |
Czech Republic |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE26725 |
Gene expression analysis of 12 B-cell Chronic Lymphocytic Leukemia samples and 5 CD19+ control samples |
|
