|
| Status |
Public on Dec 05, 2016 |
| Title |
E16 ipsi RGCs, biological rep2 |
| Sample type |
RNA |
| |
|
| Source name |
ipsilateral RGCs isolated from E16 mouse embryos
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL/6J
age: E16 embryos
tissue: embryonic retina
|
| Treatment protocol |
E16.5 embryos were removed from pregnant females under anesthesia and decapitated in DMEM/F12 buffer over ice. The head was immersed in DMEM and hard palate and skull base were removed to expose the optic chiasm and tract. Tetramethylrhodamine-conjugated dextran MW3000 was applied to the severed optic tract. Heads were incubated in pre-oxygenated bubbling ACSF at room temperature for 2 hours followed by immediate harvesting of retinas by dissection. Pooled ipsilateral or contralateral retinas from 2 litters were dissociated using papain. Rhodamine-positive neurons, with exclusion of DAPI-stained dead cells were collected by FACS.
|
| Growth protocol |
E16.5 were harvested from pregnant females housed in a barrier facility in a timed pregnancy colony. Conception is assumed to take place at approximately midnight, and E0.5 refers to the day on which the vaginal plug was detected.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA purification was performed immediately following FACS using the Absolutely RNA Nanoprep kit (Stratagene) according to the manufacturer's instructions
|
| Label |
biotin
|
| Label protocol |
Amplified cDNA was generated from purified RNA using Ovation Pico WTA System (Nugen) of which 5μg was then labeled and fragmented using the Encore Biotin Module (Nugen) according to the manufacturer's instructions.
|
| |
|
| Hybridization protocol |
Labeled cDNA was hybridized on Mouse Genome 430 2.0 Array chips Mouse Genome 430 2.0 Array chips
|
| Scan protocol |
Arrays were scanned according to the Affymetrix protocol
|
| Description |
RGCs retrogradely labeled from optic tract post-optic chiasm decussation
|
| Data processing |
Affymetrix Expression Console (1.1) was used for RMA normalization of all microarray data
|
| |
|
| Submission date |
Jun 16, 2016 |
| Last update date |
Dec 05, 2016 |
| Contact name |
Qing Wang |
| Organization name |
Columbia University
|
| Department |
Pathology and Cell Biology
|
| Lab |
Carol Ann Mason
|
| Street address |
P&S Bldg 14-509, 630 W 168th St
|
| City |
New York |
| State/province |
NY |
| ZIP/Postal code |
10032 |
| Country |
USA |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
| GSE83461 |
Ipsilateral and contralateral retinal ganglion cells express distinct genes during decussation at the optic chiasm |
|
