|
| Status |
Public on Oct 31, 2014 |
| Title |
single cell profile of retinal Muller Glial Cell 25 weeks Rhodko A3 |
| Sample type |
RNA |
| |
|
| Source name |
Rhod-ko single retinal Muller Glial Cell, 25 wks
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57/BL6
genotype/variation: Rhod-ko
tissue: retina
developmental stage: 25 wks
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Retinas were dissociated with papain, single Muller Glial cells were isolated with a micropipette based on their distinct morphology and expelled into cold lysis buffer (10 mM Tris-HCl [pH 8.3], 50 mM KCl, 1.5 mM MgCl2, 5 mM DTT, 0.5% NP-40). Total mRNAs were reverse transcribed using Superscript II (Invitrogen) in combination with an oligo dT primer. CDNAs were tailed with poly As using terminal deoxynucleotidyl transferase (TdT) and PCR amplified for 35 cycles using the same oligo dT primer.
|
| Label |
biotin
|
| Label protocol |
Ten micrograms of each single cell cDNA was digested with Dnase I for 13 minutes at 37C. Heated to 99C for 15 minutes and biotin labeled using Biotin N6 ddATP (Enzo Biosciences) and TdT (Roche) at 37C for 1.5 hours.
|
| |
|
| Hybridization protocol |
Affymetrix
|
| Scan protocol |
Affymetrix, Genechip Affymetrix Scanner 7G
|
| Description |
Rhodopsin knockout (Rhod-ko) mice are deleted in the rod-specific Rhodopsin gene
|
| Data processing |
The Affymetrix MAS5.0 algorithm was used to normalize data.
|
| |
|
| Submission date |
Jan 27, 2012 |
| Last update date |
Oct 31, 2014 |
| Contact name |
Karin Roesch |
| Organization name |
Harvard Medical School
|
| Street address |
77 Avenue Louis Pasteur
|
| City |
Boston |
| State/province |
MA |
| ZIP/Postal code |
02115 |
| Country |
USA |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
| GSE35386 |
GENE EXPRESSION CHANGES WITHIN MÜLLER GLIAL CELLS DURING RETINITIS PIGMENTOSA |
|
