|
| Status |
Public on Jun 30, 2011 |
| Title |
CPG-stimulated B-CLL repl 15 |
| Sample type |
RNA |
| |
|
| 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 using the TRIZOL Reagent.
|
| Label |
Cy3
|
| Label protocol |
Samples were labeled with Cy3 dye using the miRNA Complete Labeling System & Hyb Kit (Agilent Technologies)
|
| |
|
| Hybridization protocol |
The fluorescent labeled samples were hybridized to the Human miRNA microarray Version 3 from the Sanger database v12.0 (Agilent Technologies)
|
| Scan protocol |
The microarray slides were analyzed with an Agilent Microarray Scanner.
|
| Description |
G220_CpG
|
| Data processing |
The hybridization signal values for the multiple probes for each miRNA were obtain with the use of Agilent Feature Extraction Software 10.7.3 (Agilent Technologies). The data were analyzed by R and limma packages. The the signal is background corrected by fitting a normal-exponential convolution model to a vector of observed intensities. The signal is normalized between arrays using the quantile method.
|
| |
|
| Submission date |
Jun 20, 2011 |
| Last update date |
Nov 15, 2012 |
| 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 |
GPL14767 |
| Series (2) |
| GSE30106 |
miRNA expression profile of unmutated and mutated CLL cells |
| GSE30107 |
miRNA and mRNA expression profile of CLL cells |
|
