|
Status |
Public on Aug 18, 2011 |
Title |
CPG-stimulated B-CLL_mutated repl 7 |
Sample type |
RNA |
|
|
Channel 1 |
Source name |
CPG stimulated purified cell of CLL mutated
|
Organism |
Homo sapiens |
Characteristics |
cell type: purified CLL cells
|
Extracted molecule |
total RNA |
Extraction protocol |
CLL cells were purified by negative selection using anti-CD3, anti-CD14 and anti-CD16 mouse monoclonal antibodies and Dynabeads coated with a pan anti-mouse IgG antibody. The purity of the CLL cells after negative selection was monitored by flow-cytometry and the percentage of CD5+/CD19+ cells exceeded 98% for each sample. CLL cells were resuspended in RPMI complete medium at a density of 2 × 105 and stimulated with 7.5 μg/ml complete phosphorothioate CpG ODN oligonucleotide 2006 (5′-TCGTCGTTTTGTCGTTTTGTCGTT-3′) or left unstimulated for 18 h.Total RNA was extracted according to TRIZOL reagent protocol
|
Label |
Cy5
|
Label protocol |
Total RNA was labelled according to Ambion protocol. 825 ng of Cy3-labeled reference aRNA (according to Agilent protocol)
|
|
|
Channel 2 |
Source name |
reference
|
Organism |
Homo sapiens |
Characteristics |
sample origin: pooled normal PB B cells of healthy donors
|
Extracted molecule |
total RNA |
Extraction protocol |
CLL cells were purified by negative selection using anti-CD3, anti-CD14 and anti-CD16 mouse monoclonal antibodies and Dynabeads coated with a pan anti-mouse IgG antibody. The purity of the CLL cells after negative selection was monitored by flow-cytometry and the percentage of CD5+/CD19+ cells exceeded 98% for each sample. CLL cells were resuspended in RPMI complete medium at a density of 2 × 105 and stimulated with 7.5 μg/ml complete phosphorothioate CpG ODN oligonucleotide 2006 (5′-TCGTCGTTTTGTCGTTTTGTCGTT-3′) or left unstimulated for 18 h.Total RNA was extracted according to TRIZOL reagent protocol
|
Label |
Cy3
|
Label protocol |
Total RNA was labelled according to Ambion protocol. 825 ng of Cy3-labeled reference aRNA (according to Agilent protocol)
|
|
|
|
Hybridization protocol |
The Hybridization was performed acording to Ambion protocol
|
Scan protocol |
Slides were analysed by Agilent Microarray Scanner G2565AA
|
Description |
Biological replicate 7 of 8 251485057174_1_1_G202_CpG
|
Data processing |
Background subtraction method: Marginal log-likelihood of foreground values for normal + exponential model and its derivatives.Whithin Normalization method: loess. Between Normalization method: quantile
|
|
|
Submission date |
Aug 12, 2011 |
Last update date |
Aug 18, 2011 |
Contact name |
daniela marconi |
E-mail(s) |
[email protected]
|
Organization name |
CRO AVIANO
|
Street address |
Via franco Gallini 2
|
City |
Aviano |
ZIP/Postal code |
33081 |
Country |
Italy |
|
|
Platform ID |
GPL4133 |
Series (2) |
GSE30107 |
miRNA and mRNA expression profile of CLL cells |
GSE31360 |
CPG stimulation for Gene Expression of mutated B-CLL |
|