|
| Status |
Public on Jun 01, 2011 |
| Title |
Braak I-II, APOE e4+ sample 3 |
| Sample type |
RNA |
| |
|
| Source name |
JS3
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: temporal cortex
cell type: astrocytes
braak stage: Braak I-II
apoe status: APOE e4+
|
| Treatment protocol |
7um frozen temporal cortex sections were fixed in acetone and immunostained for GFAP using a rapid staining protocol. They were then washed and dehydrated through graded ethanol concentrations (70%, 95%, 100%) and cleared in xylene.
From each individual case, approximately 1500 astrocytes were isolated using a PixCell II laser-capture microdissection system (Arcturus Bioscience)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
PicoPure RNA Extraction Kit was used for RNA extraction according to manufacturers' instructions
|
| Label |
biotin
|
| Label protocol |
Biotinylation of antisense RNA carried out according to Affymetrix protocol
|
| |
|
| Hybridization protocol |
7µg of cRNA molecules were heat fragmented and applied to the Human Genome HGU133 Plus 2.0 GeneChip in a hybridisation solution according to the Affymetrix protocol. Hybridisation took place overnight in a rotating hybridisation platform at 45°C for 16 hours. Post hybridisation stringency washing was done using the Affymetrix fluidics station again following the protocol outlined in the Affymetrix instructions
|
| Scan protocol |
the GeneChips were scanned in the Affymetrix GCS3000 scanner and the resultant image processed using the GCOS software package to prepare the .CEL files
|
| Description |
Gene expression data from human astrocytes
|
| Data processing |
The data were using Array Assist 3 software
|
| |
|
| Submission date |
Jun 01, 2011 |
| Last update date |
Jun 01, 2011 |
| Contact name |
Paul Roy Heath |
| E-mail(s) |
[email protected]
|
| 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 |
GPL570 |
| Series (1) |
| GSE29652 |
Microarray analysis of the astrocyte transcriptome in the ageing brain: relationship to Alzheimer's pathology and ApoE genotype |
|
