|
| Status |
Public on Feb 01, 2011 |
| Title |
PGC_bFGF replacement_rep2 |
| Sample type |
RNA |
| |
|
| Source name |
PGCs after bFGF replacement
|
| Organism |
Gallus gallus |
| Characteristics |
breed: Korean Oge (KO)
tissue: whole blood
cell type: primordial germ cells (PGCs)
age: embryonic day 2.5
treatment: basic fibroblast growth factor (bFGF) replacement
|
| Treatment protocol |
Cells were collected before and after withdrawal of bFGF for 24h, and 24h after bFGF replacement.
|
| Growth protocol |
Approximately 2 μl of whole blood cells was taken from the dorsal aortae of stage 14–15 (H&H) (50–54 h of incubation) Korean Oge chicken embryos and cultured in medium comprising knockout Dulbecco’s modified Eagle’s medium (Invitrogen, Carlsbad, CA), 7.5% fetal bovine serum (Hyclone, Logan, UT), 2.5% chicken serum (Sigma-Aldrich, St. Louis, MO), 2 mM GlutaMAX-I Supplement (Invitrogen), 1× nucleosides (Millipore, Temecula, CA), 1× nonessential amino acids, β-mercaptoethanol, and 10 ng·ml–1 human bFGF (Sigma-Aldrich).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from three different treated cell samples using a RNeasy Total RNA Isolation Kit (Qiagen).
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 6 ug total RNA (Expression Analysis Technical Manual, 2001, Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 ug of cRNA were hybridized for 16 hr at 45C on GeneChip Chicken Genome Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the Hewlett-Packard GeneArray Scanner G2500A.
|
| Description |
Gene expression data from PGCs after bFGF replacement.
|
| Data processing |
The data were analyzed with Microarray Suite version 5.0 (MAS 5.0) using Affymetrix default analysis settings and global scaling as normalization method. The trimmed mean target intensity of each array was arbitrarily set to 100.
|
| |
|
| Submission date |
Jun 27, 2010 |
| Last update date |
Feb 01, 2011 |
| Contact name |
JAE YONG HAN |
| E-mail(s) |
jaehan@snu.ac.kr
|
| Phone |
82-2-880-4810
|
| Organization name |
Seoul National University
|
| Department |
Department of Agricultural Biotechnology
|
| Lab |
Animal Genetic Engineering
|
| Street address |
1 Gwanak-ro, Gwanak-gu
|
| City |
Seoul |
| ZIP/Postal code |
08826 |
| Country |
South Korea |
| |
|
| Platform ID |
GPL3213 |
| Series (1) |
| GSE22592 |
Expression data from cultured chicken primordial germ cells (PGCs) |
|
