|
| Status |
Public on Dec 02, 2009 |
| Title |
OM-OECs vs. OB-OECs, 3 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Olfactory mucosa OECs
|
| Organism |
Rattus norvegicus |
| Characteristics |
strain: Fischer
gender: male
tissue: olfactory mucosa
cell type: olfactory ensheathing cells
|
| Growth protocol |
OM-OECs and OB-OECs were cultivated with the same protocol (A. Ramon-Cueto and M. Nieto-Sampedro, 1992).
|
| Extracted molecule |
polyA RNA |
| Extraction protocol |
Cell samples were lysed using SuperAmp Lysis Buffer and stored appropriately according to the instructions of the SuperAmp Preparation Kit (Miltenyi Biotec). mRNA was isolated via magnetic bead technology.
|
| Label |
Cy5
|
| Label protocol |
SuperAmp RNA amplification was performed according to Miltenyi Biotec's undisclosed procedure. The amplification is based on a global PCR protocol using mRNA-derived cDNA. 250 ng of each of the cDNAs were used as template for Cy3 and Cy5 labeling which was performed according to Miltenyi Biotec's undisclosed protocol. OM-OECs are labeled with Cy5 and OB-OECs are labeled with Cy3.
|
| |
|
| Channel 2 |
| Source name |
Olfactory bulb OECs
|
| Organism |
Rattus norvegicus |
| Characteristics |
strain: Fischer
gender: male
tissue: olfactory bulb
cell type: olfactory ensheathing cells
|
| Growth protocol |
OM-OECs and OB-OECs were cultivated with the same protocol (A. Ramon-Cueto and M. Nieto-Sampedro, 1992).
|
| Extracted molecule |
polyA RNA |
| Extraction protocol |
Cell samples were lysed using SuperAmp Lysis Buffer and stored appropriately according to the instructions of the SuperAmp Preparation Kit (Miltenyi Biotec). mRNA was isolated via magnetic bead technology.
|
| Label |
Cy3
|
| Label protocol |
SuperAmp RNA amplification was performed according to Miltenyi Biotec's undisclosed procedure. The amplification is based on a global PCR protocol using mRNA-derived cDNA. 250 ng of each of the cDNAs were used as template for Cy3 and Cy5 labeling which was performed according to Miltenyi Biotec's undisclosed protocol. OM-OECs are labeled with Cy5 and OB-OECs are labeled with Cy3.
|
| |
|
| |
| Hybridization protocol |
The corresponding Cy3- and Cy5-labeled cDNAs were combined and hybridized overnight (17 hours, 65°C) to Agilent Whole Rat Genome Oligo Microarrays 4x44K using Agilent's recommended hybridization chamber and oven.
|
| Scan protocol |
Agilent's DNA microarray scanner (Agilent Technologies).
|
| Description |
Biological replicate 3 of 4.
|
| Data processing |
Images were quantified using Agilent Feature Extraction Software (version 9.1). Agilent Feature Extraction Software was used for background subtraction and Linear LOWESS normalization.
|
| |
|
| Submission date |
Dec 01, 2009 |
| Last update date |
Dec 01, 2009 |
| Contact name |
nicolas guerout |
| E-mail(s) |
nicolas_guerout_76@hotmail.com
|
| Phone |
0235148390
|
| Fax |
0235148388
|
| Organization name |
Faculty of medecine of Rouen
|
| Department |
EA3830
|
| Lab |
Experimental Surgery
|
| Street address |
22 Bd Gambetta
|
| City |
Rouen |
| State/province |
Normandy |
| ZIP/Postal code |
76000 |
| Country |
France |
| |
|
| Platform ID |
GPL4135 |
| Series (1) |
| GSE19250 |
Comparative Gene Expression Profiling of Olfactory Ensheathing Cells from Olfactory Bulb and Olfactory Mucosa |
|
