|
| Status |
Public on Feb 15, 2014 |
| Title |
Gene expression of myoblasts before myogenic stimuli |
| Sample type |
RNA |
| |
|
| Source name |
myoblasts
|
| Organism |
Homo sapiens |
| Characteristics |
gender: male
age: 7
ethnicity: Caucasian
|
| Treatment protocol |
Differentiation to myotubes were stimulated by switching the culture medium to DMEM-F12 (Thermo scientific) with 2% horse serum (Sigma Aldrich) when cells were reachd about 50% confluent. Medium was replaced every other day, and changed to SkGM-2 medium at day 8. SkGM-2 medium was replaced every other day until day 15. Cells were collected on day 1, day 3, day 8 and day 15.
|
| Growth protocol |
Myoblasts were cultured with SkGM-2 medium (Lonza) at 37C 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
ISOGEN (Nippon gene) extraction of total RNA was performed according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the Affymetrix Gene Profiling Regent User Guide (Affymetrix) from 200 ng of total RNA.
|
| |
|
| Hybridization protocol |
Following fragmentation, 20 ug of cRNA were hybridized for 16 hr at 45C on Affymetrix Human Genome U133 Plus 2.0 Arrays (Affymetrix). GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the GeneChip scanner 3000 7G.
|
| 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 |
Feb 14, 2014 |
| Last update date |
Feb 15, 2014 |
| Contact name |
Kohei Miyata |
| E-mail(s) |
md100029@cis.fukuoka-u.ac.jp
|
| Phone |
+81928011011
|
| Organization name |
Fukuoka University
|
| Department |
Obstetrics and Gynecology
|
| Street address |
7-45-1 Nanakuma, Jounannku
|
| City |
Fukuoka-city |
| State/province |
Fukuoka |
| ZIP/Postal code |
8140131 |
| Country |
Japan |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE55034 |
DNA methylation and gene expression analysis during myogenic differentiation |
|
