|
| Status |
Public on Jun 30, 2011 |
| Title |
CPG-stimulated B-CLL repl 8 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
CPG stimulated purified cell of CLL
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: B-CLL
|
| Treatment 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.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
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
|
| Treatment 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.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
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 8 of 9
|
| 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 |
Jun 20, 2011 |
| Last update date |
Jun 30, 2011 |
| Contact name |
daniela marconi |
| E-mail(s) |
daniela.marconi@gmail.com
|
| Organization name |
CRO AVIANO
|
| Street address |
Via franco Gallini 2
|
| City |
Aviano |
| ZIP/Postal code |
33081 |
| Country |
Italy |
| |
|
| Platform ID |
GPL4133 |
| Series (2) |
| GSE30105 |
CPG stimulation for Gene Expression of unmutated B-CLL |
| GSE30107 |
miRNA and mRNA expression profile of CLL cells |
|
