|
| Status |
Public on Apr 10, 2017 |
| Title |
MN innervating EDL [M33EDL] |
| Sample type |
RNA |
| |
|
| Source name |
Laser captured motor neurons
|
| Organism |
Mus musculus |
| Characteristics |
genotype: Wild type
strain: FVB
tissue: Laser captured motor neurons innervating extensor digitorum longus muscle
|
| Extracted molecule |
total RNA |
| Extraction protocol |
20 um cervical spinal cord frozen sections on PEN membrane slides were acetone fixed and dehydrated through graded ethanol concentrations (70%, 90%, 100%). Motor neurons with retrogradely transported WGA-HRP were stained with NovaRed peroxidase (Vector Labs).
From each individual all stained motor neurons (MN) were isolated using a Veritas laser-capture microdissection system (Applied Biosystems)
PicoPure RNA Extraction Kit was used for RNA extraction according to manufacturers' instructions
|
| Label |
biotin
|
| Label protocol |
Biotinylation of sense DNA strand was carried out according to Affymetrix protocol
|
| |
|
| Hybridization protocol |
5.5µg of sense DNA molecules were heat fragmented and applied to the Mouse transcriptome array 1 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 |
Scanned on an Affymetrix® GeneChip Scanner 3000 according to manufacturer's protocols
|
| Description |
Laser captured motor neurons innervating extensor digitorum longus muscle
|
| Data processing |
Data were RMA normalised using the Affymetrix transcriptome analysis console package
|
| |
|
| Submission date |
Sep 14, 2016 |
| Last update date |
Apr 10, 2017 |
| 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 |
GPL20775 |
| Series (1) |
| GSE86908 |
Bioenergetics status determines motor neuron vulnerability in spinal muscular atrophy |
|
