|
| Status |
Public on Apr 01, 2011 |
| Title |
blood_day0_patient5 |
| Sample type |
RNA |
| |
|
| Source name |
Blood
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: blood
patient: 5
gender: male
age: 38
|
| Treatment protocol |
Blood samples were collected with PAXgene Blood RNA tubes
|
| Growth protocol |
Peripheral blood cells
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA extraction was performed with PAXgene Blood RNA kit according to manufactors instructions, and RNA quality assesed with Bioanalyzer
|
| Label |
biotin
|
| Label protocol |
Isolated total RNA was used with GeneChip WT cDNA synthesis and amplification kit (Affymetrix#900672) to produce fragmented single-stranded DNA, which was labeled using terminal labeling kit (Affymetrix#900670) to generate biotin-labeled sense targeted DNA probes.
|
| |
|
| Hybridization protocol |
Fragmented and labeled DNA (2500ng) was used in 100 µl hybridization cocktail containing control oligonucleotide B2 (50pM) and hybridization controls (Affymetrix #900454), of which 80 µl was hybridised to each chip for 17h in the hybridization oven at 45ºC and rotating at 60rpm. Arrays were processed using Genechip Fluidics Station 450 according to recommended protocols (FS450_007) of double-staining and post-hybridization washes using GeneChip hybridization, wash and stain kit (Affymetrix #900720).
|
| Scan protocol |
Fluorescent images were captured using gene Array scanner 3000 7G and Affymetrix GeneChip Command Console, v 3.1 software (Affymetrix).
|
| Data processing |
Raw data were processed using Partek Genomics Suite v. 6.5 to generate RMA normalised intensities. Differential expression was assessed using One-way or Two-way ANOVA in the Partek software dependent of if the samples were paired or not.
|
| |
|
| Submission date |
Feb 08, 2011 |
| Last update date |
Apr 01, 2011 |
| Contact name |
Jan-Erik Berdal |
| E-mail(s) |
[email protected]
|
| Phone |
004748205817
|
| Organization name |
AHUS
|
| Department |
medical department
|
| Street address |
Sykehusvn.
|
| City |
Lørenskog |
| State/province |
Akershus |
| ZIP/Postal code |
1474 |
| Country |
Norway |
| |
|
| Platform ID |
GPL6244 |
| Series (1) |
| GSE27131 |
Peripheral blood cells expression data form 7 patients with severe pdm(H1N1) influensa and 7 gender and age matched healthy controls |
|
