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Status |
Public on Nov 11, 2008 |
Title |
GE110_GWP |
Sample type |
genomic |
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Source name |
Human B-cell chronic lymphocytic leukemia patient GE110
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Organism |
Homo sapiens |
Characteristics |
Legend: VH status = IgVH mutational status (Fais et al. J Clin Invest, 1998) Sex: F; VH status: mut
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Treatment protocol |
Peripheral blood mononuclear cells from B-CLL patients were isolated by Ficoll-Hypaque (Seromed, Biochrom KG, Berlin, Germany) density-gradient centrifugation and the proportion of CD5/CD19/CD23 triple positive B cells in the suspension was determined by direct immunofluorescence performed using a FACS-sort flow cytometer (Becton Dickinson & Co, Sunnyvale, CA) with antibodies to: CD19 FITC/PE, CD23 PE and CD5 Cy-Chrome (Becton Dickinson). If B-CLL cells were less than 90%, T cells, NK cells and monocytes were removed by negative selection using CD3, CD56, CD16, and CD14 monoclonal antibody (mAb) treatment (Becton Dickinson) followed by magnetic beads (Goat Anti-Mouse IgG Dynabeads, Dynal Biotech ASA, Oslo, Norway).
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Extracted molecule |
genomic DNA |
Extraction protocol |
Genomic DNA extraction was performed using Wizard® Genomic DNA Purification kit according to the manufacturer's instructions (Promega).
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Label |
biotin
|
Label protocol |
Biotinylated DNA were prepared according to the standard Affymetrix protocol starting from 250 nanograms of genomic DNA.
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Hybridization protocol |
Following fragmentation, 40 micrograms of biotin-labeled DNA were hybridized for 16 hr at 48°C on GeneChip Human Mapping 50K XbaI Arrays. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
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Scan protocol |
Human Mapping 50K XbaI arrays were scanned using the GeneChip Scanner 3000 7G (Affymetrix).
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Description |
Genome-wide profiling data from human B-cell chronic lymphocytic leukemia patient GE110
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Data processing |
The array image was acquired using Affymetrix GeneChip® Operating Software (GCOS version 1.4). Copy number values for individual SNPs were extracted and converted from CEL files into signal intensities using GTYPE 4.1 and Affymetrix Copy Number Analysis Tool (CNAT 4.0.1) softwares. Genomic Smoothing analysis was performed by using the smoothing window of 1 Mb, and inferred copy number states were derived from a Hidden Markov Model (HMM) based algorithm implemented in CNAT 4.0.1.
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Submission date |
Apr 03, 2008 |
Last update date |
Nov 11, 2008 |
Contact name |
Luca Agnelli |
E-mail(s) |
luca.agnelli@istitutotumori.mi.it, luca.agnelli@gmail.com
|
Phone |
+390223903581
|
Organization name |
IRCCS Istituto Nazionale dei Tumori
|
Department |
Department of Advanced Diagnostics
|
Street address |
Venezian 1
|
City |
MILAN |
ZIP/Postal code |
20133 |
Country |
Italy |
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Platform ID |
GPL2005 |
Series (2) |
GSE11036 |
Molecular and transcriptional characterization of chromosome 17p loss in chronic lymphocytic leukemia, experiment B |
GSE11038 |
Molecular and transcriptional characterization of chromosome 17p loss in chronic lymphocytic leukemia |
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