|
| Status |
Public on Jun 01, 2014 |
| Title |
ALS with SOD1 mutation patient 28 |
| Sample type |
RNA |
| |
|
| Source name |
cultured fibroblasts
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: fibroblasts
patient group: ALS due to mtSOD1
|
| Growth protocol |
Monolayers of fibroblasts were then grown in T75 flasks with 20mL of Ham’s F-10 Media (Gibco) with 10% Fetal Calf serum (Biosera), 1% Penicillin-streptomycin antibiotic solution, 0.25% Holo-transferrin (Sigma) and 2.5 ng epidermal growth factor (Sigma).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was extracted from the fibroblasts using the Qiagen RNeasy Mini Kit (Qiagen).
|
| Label |
biotin
|
| Label protocol |
The Ambion WT Expression Kit was used to amplify the RNA and convert it to cDNA. The cDNA was then fragmented and biotin-labelled using the Affymetrix GeneChip WT Terminal Labelling Kit
|
| |
|
| Hybridization protocol |
hybridized to GeneChip Human Exon 1.0 ST Arrays, washed and stained using the Affymetrix GeneChip Hybridization, Wash and Stain Kit and an Affymetrix Fluidics Station 450
|
| Scan protocol |
scanned on an Affymetrix® GeneChip Scanner 3000 according to manufacturer's protocols
|
| Description |
Fibroblasts from individuals with SOD1 mutations
|
| Data processing |
The Partek Genomics Suite was used to normalize (by GC RMA) and then analyse the microarray data following Affymetrix guidelines. Core probesets only were used.
|
| |
|
| Submission date |
Nov 21, 2011 |
| Last update date |
Jun 01, 2014 |
| Contact name |
Paul Roy Heath |
| E-mail(s) |
p.heath@sheffield.ac.uk
|
| Phone |
+44 114 2222254
|
| Organization name |
University of Sheffield
|
| Department |
Academic Neurology Unit
|
| Lab |
Heath
|
| Street address |
SITraN, 385a Glossop Road
|
| City |
Sheffield |
| State/province |
S. Yorks |
| ZIP/Postal code |
S10 2HQ |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL5188 |
| Series (2) |
| GSE33855 |
Loss of nuclear TDP-43 in ALS causes altered expression of splicing machinery and widespread dysregulation of RNA splicing in motor neurons [fibroblasts] |
| GSE56504 |
Loss of nuclear TDP-43 in ALS causes altered expression of splicing machinery and widespread dysregulation of RNA splicing in motor neurons |
|
