|
| Status |
Public on Oct 03, 2013 |
| Title |
adipose- derived stem cells Donor 2 |
| Sample type |
RNA |
| |
|
| Source name |
Adult stem cell
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: Adult stem cell
|
| Extracted molecule |
total RNA |
| Extraction protocol |
For the expression microarray experiment in hASCs and in vitro differentiated cells, total RNA was prepared using TRIZOL® (Invitrogen, Carlsbad, CA) and further purified using RNeasy columns (Qiagen, GmbH) according to the manufacturer’s instructions. The integrity of RNA was monitored by both Byoanalyzer and spectrometric measurement
|
| Label |
Biotin
|
| Label protocol |
Biotinylated target RNA was prepared from 200 ng of total RNA using the Affymetrix protocol. Briefly, double-stranded cDNA was prepared from the RNA template using the Ambion WT Expression kit following manufacturer’s instructions.
|
| |
|
| Hybridization protocol |
The resulting cRNA target was randomly fragmented using UDG (Uracil DNA Glycosylase) and APE1 (apurinic/apyrimidic endonuclease 1), labelled with the terminal transferase (WT Terminal Labelling kit, Affymetrix) and hybridized on the Human Gene 1.1 ST 16-array (Affymetrix, Santa Clara, CA)
|
| Scan protocol |
The hybridization reactions were processed and scanned according to the standard GeneTitanTM (Affymetrix, Santa Clara, CA) protocols.
|
| Description |
ASC2
|
| Data processing |
All arrays were globally scaled to a target-intensity value of 100 and then the scaling factor, background, noise, and percentage present were calculated according to the Command Console (AGCC 2.0) and Expression Console (EE 1.1) softwares (Affymetrix, Santa Clara, CA). All resulting datasets were filtered using the absolute call metric (present or absent) using Microsoft Access (Microsoft Corporation, Redmond, WA). The resulting values represent the transcript abundance (absolute value) following Affymetrix statistical’s procedure. Three biological replicates were done for each ASC sample and in vitro differentiated cells were analysed in duplicate.
|
| |
|
| Submission date |
Apr 17, 2012 |
| Last update date |
Oct 04, 2013 |
| Contact name |
Manel Esteller |
| Organization name |
IDIBELL
|
| Department |
PEBC
|
| Lab |
Cancer Epigenetics
|
| Street address |
Hospital Duran i Reynals Av. Gran Via s/n km, 2.7
|
| City |
L'Hospitalet de Llobregat |
| State/province |
Barcelona |
| ZIP/Postal code |
08908 |
| Country |
Spain |
| |
|
| Platform ID |
GPL11532 |
| Series (1) |
| GSE37329 |
Genome-wide gene expression during osteogenic and myogenic differentiation from adipose- derived stem cells. |
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