|
| Status |
Public on May 05, 2014 |
| Title |
Human ADSC_sample 1 |
| Sample type |
RNA |
| |
|
| Source name |
Human ADSC-1, Passage 5
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: Adipose
|
| Growth protocol |
All cell lines were cultured in Dulbecco’s modified Eagle’s medium (Gibco) supplemented with 10% heat-inactivated fetal bovine serum (Gibco), 100 units/ml penicillin, and 100 µg/ml streptomycin (Gibco) in 5% CO2 in a humidified incubator at 37°C. All hMSC experiments were performed at passage 3 to 6.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated using Trizol (Life Technologies) according to the manufacturer's instructions.
|
| Label |
Cy3
|
| Label protocol |
For control and test RNAs, the synthesis of target miRNA probes and hybridization were performed using Agilent’s miRNA Labeling Reagent and Hybridization kit (Agilent Technology, USA) according to the manufacturer’s instructions. Briefly, each 100 ng of total RNA were dephosphorylated with ~15 Units of calf intestine alkaline phosphatase (CIP), followed by RNA denaturation with ~40% DMSO and 10 min incubation at 100 °C. Dephosphorylated RNA were ligated with pCp-Cy3 mononucleotide and purified with MicroBioSpin 6 columns (Bio-rad, USA).
|
| |
|
| Hybridization protocol |
After purification, labeled samples were resuspended with Gene Expression blocking Reagent and Hi-RPM Hybridization buffer, followed by boiling for 5 min at 100 °C and 5 min chilled on ice. Finally, denatured labeled probes were pipetted onto assembled Agilent’s miRNA Microarray and hybridized for 20 hours at 55°C with 20 RPM rotating in Agilent Hybridization oven (Agilent Technology, USA).
|
| Scan protocol |
The hybridized images were scanned using Agilent’s DNA microarray scanner and quantified with Feature Extraction Software (Agilent Technology, Palo Alto, CA).
|
| Description |
miRNA
|
| Data processing |
All data normalization and further analysis were performed using GeneSpringGX 7.3 (Agilent Technology, USA).
|
| |
|
| Submission date |
May 04, 2014 |
| Last update date |
May 05, 2014 |
| Contact name |
Nayoung Suh |
| Organization name |
Asan Medical Center, University of Ulsan College of Medicine
|
| Department |
Department of Medicine
|
| Lab |
Asan Institute for Life Sciences
|
| Street address |
88 Olympic-ro 43-gil, Songpagu
|
| City |
Seoul |
| ZIP/Postal code |
138-736 |
| Country |
South Korea |
| |
|
| Platform ID |
GPL15019 |
| Series (1) |
| GSE57266 |
Human Mesenchymal Stem Cells (hMSCs): Fibroblast vs. hMSCs from various origins |
|
