|
| Status |
Public on Jul 31, 2007 |
| Title |
SympatheticNeuron_WithMyocyteTarget |
| Sample type |
RNA |
| |
|
| Source name |
Sympathetic Neuron cultured with Myocyte Target
|
| Organism |
Rattus norvegicus |
| Characteristics |
Sympathetic neuron culture from neonatal superior cervical ganglia of Simonson Albino rat cultured with neonatal cardiac myocytes 2 days in vitro
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Neurons were isolated from cultures containing myocytes or other flat cells the culture dishes were washed with PBS and incubated with 1 mM EDTA in PBS for 2 min in 37°C. The less adherent sympathetic neurons were then removed from the plate by pipetting 1 mM EDTA in PBS over the dish. Rinsed neurons were collected and centrifuged in a clinical centrifuge at 3,000 rpm for 10 min. Total RNA was extracted using the RNeasy mini kit (Qiagen) according to manufacture's protocol.
|
| Label |
biotin
|
| Label protocol |
The synthesis of biotin-labeled cRNA was performed using the BioArray High-Yield Transcript Labeling Kit (Enzo Biochem, New York, NY).
|
| |
|
| Hybridization protocol |
Hybridization was done according to Affymetrix protocol for 16 hours.
Staining and Washing was also done according to Affymetrix protocol (Fluidics Scripts: EukGE-WS2v4_450).
|
| Scan protocol |
Scan was done by Affymetrix GeneChip Scanner 3000 according to Affymetrix protocol
|
| Description |
Affymetrix Rat Expression Set 230A oligonucleotide arrays with 15,900 probe sets were screened with probes derived from neurons grown alone. Total RNA was extracted using the RNeasy mini kit (Qiagen). Eukaryotic probe preparation, hybridization, array washing, staining and scanning were performed according to Affymetrix manuals (Rev. 5).
|
| Data processing |
Affymetrix MAS5.0 algorithm was used for data processing
|
| |
|
| Submission date |
Jun 29, 2007 |
| Last update date |
Aug 14, 2011 |
| Contact name |
Susan J Birren |
| E-mail(s) |
birren@brandeis.edu
|
| Organization name |
Brandeis University
|
| Department |
Biology
|
| Lab |
Birren
|
| Street address |
415 South st
|
| City |
Waltham |
| State/province |
MA |
| ZIP/Postal code |
02454 |
| Country |
USA |
| |
|
| Platform ID |
GPL341 |
| Series (1) |
| GSE8393 |
Patterns of gene expression following contact of sympathetic neurons with their myocyte targets |
|
