|
| Status |
Public on Dec 13, 2014 |
| Title |
Subject3_day0 |
| Sample type |
RNA |
| |
|
| Source name |
Skeletal muscle cells_proliferating (myoblasts)
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: vastus lateralis muscle
cell type: Human skeletal muscle cells
time point: before induction of differentiation (proliferating myoblasts)
|
| Growth protocol |
Human skeletal muscle cells (extracted from vastus lateralis muscle) was cultured in Dulbecco's Modified Eagle Medium (DMEM):Nutrient Mixture F-12 supplemented with 20% FBS, 1% fungizone and 1% Penicillin-Streptomycin. Cultures were subcultured with trypsin before reaching 70% confluence. Differentiation of myoblasts into myotubes were induced by switching to differentiation medium (DMEM (low glucose) supplemented with 2% FBS, 1% PeSt and 1% Fungizone) when cells were 80% confluent. Cultures were incubated at 37°C in 7.5% CO2 humidified chambers.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Extraction of total RNA by TRIzol (Invitrogen). Modifications of manufacturer's protocol include addition of 0.75 ml isopropanol (compared to 0.5 ml) containing glycogen with 20 minutes incubation at -20°C to precipitate RNA.
|
| Label |
Biotin
|
| Label protocol |
Sense strand cDNA was synthesized from total RNA with Ambion® WT Expression Kit (Life Techniologies) and biotin labeled with the Affymetrix® GeneChip WT Terminal Labeling Kit (Affymetrix).
|
| |
|
| Hybridization protocol |
Hybridized to Affymetrix Human Gene 1.0 ST Array according to manufacturer's protocols in a GeneChip Hybridization Oven 640 (Affymetrix). Arrays were washed and stained with a GeneChip Gluidics station 450 (Affymetrix).
|
| Scan protocol |
Arrays were scanned with a GeneChip Scanner 3000 7G (Affymetrix).
|
| Description |
Sample 3
|
| Data processing |
Pre-processing of data was performed using Robust Multi-array Average (RMA) with sketch quantile normalization with the Expression Console™ software (Affymetrix).
|
| |
|
| Submission date |
Dec 16, 2013 |
| Last update date |
Dec 13, 2014 |
| Contact name |
Anna Krook |
| E-mail(s) |
anna.krook@ki.se
|
| Organization name |
Karolinska Institutet
|
| Street address |
von Eulers väg 4a
|
| City |
Stockholm |
| ZIP/Postal code |
17177 |
| Country |
Sweden |
| |
|
| Platform ID |
GPL11532 |
| Series (2) |
| GSE53382 |
Dynamic time course mRNA profiling of human skeletal muscle cell differentiation. |
| GSE53384 |
miRNA-transcriptome expression pattern during differentiation in human skeletal muscle cells |
|
