|
| Status |
Public on May 09, 2017 |
| Title |
cell_isolated_E4_rep1 |
| Sample type |
RNA |
| |
|
| Source name |
E4 chicken retina
|
| Organism |
Gallus gallus |
| Characteristics |
tissue: dissociated retina
time point: E4
|
| Treatment protocol |
Single cells were isolated and immediately placed into cell lysis buffer.
|
| Growth protocol |
Retinas were dissected and disocciated with papain at different timepoints. There was no culture or time for growth.
|
| Extracted molecule |
polyA RNA |
| Extraction protocol |
Individual chicken retinal cells were harvested using a pulled glass pipette and were expelled into cold lysis buffer. The total single cell RNA was reverse transcribed using Superscript III combined with an oligo dT primer. This first strand cDNA product was tailed with A’s 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 |
Amplified RNA was hybridized to Affymetrix GeneChip Chicken arrays using standard Affymetrix protocols at either the GeneChip Facility at Iowa State University or the University of Iowa.
|
| Scan protocol |
Affymetrix GeneChip Chicken arrays were scanned using standard protocols at either the GeneChip Facility at Iowa State University or the University of Iowa.
|
| Description |
Gene expression data from a single developing chick retinal cell
|
| Data processing |
The data were analyzed with Microarray Suite version 5.0 (MAS 5.0) using Affymetrix default analysis settings.
|
| |
|
| Submission date |
Mar 28, 2017 |
| Last update date |
May 10, 2017 |
| Contact name |
Jeff Trimarchi |
| E-mail(s) |
jtrimarc@iastate.edu
|
| Organization name |
Iowa State University
|
| Department |
Genetics, Development and Cell Biology
|
| Street address |
2437 Pammel Drive
|
| City |
Ames |
| State/province |
IA |
| ZIP/Postal code |
50011 |
| Country |
USA |
| |
|
| Platform ID |
GPL3213 |
| Series (1) |
| GSE87663 |
Expression data from single embryonic chick retinal cells |
|
