|
| Status |
Public on Sep 05, 2014 |
| Title |
skeletal muscle tissue from healthy individual 2 |
| Sample type |
genomic |
| |
|
| Source name |
skeletal muscle
|
| Organism |
Homo sapiens |
| Characteristics |
gender: male
health status: healthy
|
| Growth protocol |
Human primary skeletal muscle myoblasts (CC-2580, Lonza) were cultured with in SkGM-2 medium (CC-3245, Lonza). Differentiation into myotubes was stimulated by switching the culture medium to DMEM-F12 (SH30023.01, Thermo scientific) supplemented with 2% horse serum (H1138, Sigma Aldrich) when cells reached about 50% confluencet (day 1). 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 days 1, day3, day8, and day15 for subsequent analyses. Human normal skeletal muscle tissues from two individuals were obtained from Tissue Solutions (SK052009, SKm1;, SKM102210A, SKm2). Human mesenchymal stem cells (hHMSCs, PT-2501, Lonza) were grown in DMEM (SH30022.01, Thermo Scientific) supplemented with 10% fetal bovinene serum (SH30070.03, Thermo Scientific). All cell lines and human tissues were derived from male individuals.
|
| Extracted molecule |
genomic DNA |
| Extraction protocol |
DNA extracted by Phenol:Chloroform:Isoamylalcohol (Sigma)
|
| Label |
Cy3 and Cy5
|
| Label protocol |
Standard Illumina labelling protocol
|
| |
|
| Hybridization protocol |
Bisulphite converted DNA was amplified, fragmented and hybridised to Illumina Infinium Human Methylation450 Beadchip using standard Illumina protocol
|
| Scan protocol |
Arrays were imaged using High Scan SQ using standard recommended Illumina scanner setting
|
| Data processing |
Normalized average beta value. The methylation level of each of the > 480,000 CpG sites was calculated using the GenomeStudio Methylation Module 1.9.0 (Illumina). Normalization was performed using Normalization to Internal Controls and Background Subtraction options implemented in the Methylation Module 1.9.0.
|
| |
|
| Submission date |
Mar 04, 2014 |
| Last update date |
Sep 05, 2014 |
| Contact name |
Kazuhiko Nakabayashi |
| E-mail(s) |
nakabaya-k@ncchd.go.jp
|
| Organization name |
National Research Institute for Child Health and Development
|
| Department |
Department of Maternal-Fetal Biology
|
| Street address |
2-10-1 Okura
|
| City |
Setagaya |
| State/province |
Tokyo |
| ZIP/Postal code |
157-8535 |
| Country |
Japan |
| |
|
| Platform ID |
GPL13534 |
| Series (1) |
| GSE55571 |
DNA methylation analysis during in vitro myogenic differentiation |
|
