|
Status |
Public on Feb 27, 2009 |
Title |
hASC-2 |
Sample type |
RNA |
|
|
Channel 1 |
Source name |
human adipose stem cells
|
Organism |
Homo sapiens |
Characteristics |
patient-derived adipose stem cells; primary cell line
|
Extracted molecule |
total RNA |
Extraction protocol |
Total RNA samples were isolated in Trizol (Invitrogen) followed by purification over a Qiagen RNeasy column (Qiagen, Valencia, CA, USA).
|
Label |
Cy5
|
Label protocol |
Using Agilent Low RNA Input Fluorescent Linear Amplification Kits, cDNA was reverse transcribed from each RNA sample, as well as from a pooled reference control, and cRNA then transcribed and fluorescently labeled with Cy5/Cy3. cRNA was purified using an RNeasy kit (Qiagen,).
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|
|
Channel 2 |
Source name |
human embryonic stem cells and embryoid bodies
|
Organism |
Homo sapiens |
Characteristics |
pooled RNA from H9 hESCs and embryoid bodies
|
Extracted molecule |
total RNA |
Extraction protocol |
Total RNA samples were isolated in Trizol (Invitrogen) followed by purification over a Qiagen RNeasy column (Qiagen, Valencia, CA, USA).
|
Label |
Cy3
|
Label protocol |
Using Agilent Low RNA Input Fluorescent Linear Amplification Kits, cDNA was reverse transcribed from each RNA sample, as well as from a pooled reference control, and cRNA then transcribed and fluorescently labeled with Cy5/Cy3. cRNA was purified using an RNeasy kit (Qiagen,).
|
|
|
|
Hybridization protocol |
825 ng of Cy3- and Cy5- labeled and amplified cRNA was hybridized to Agilent 4x44K whole human genome microarrays (G4112F) and processed according to the manufacturer’s instructions.
|
Scan protocol |
The array was scanned using an Agilent G2505B DNA microarray scanner.
|
Description |
Microarray study was performed in duplicates (hASCs, iPS cells derived from hASCs, and hESCs). Reference control was pooled RNA taken from H9 hESCs, embryoid bodies, and some differentiated cell types such as endothelial cells.
|
Data processing |
The image files were extracted using Agilent Feature Extraction software version 9.5.1 applying LOWESS background subtraction and dye-normalization.
|
|
|
Submission date |
Feb 24, 2009 |
Last update date |
Feb 26, 2009 |
Contact name |
Kitchener D. Wilson |
E-mail(s) |
kitchwilson@stanford.edu
|
Organization name |
Stanford University
|
Street address |
S140 Grant Bldg
|
City |
Stanford |
State/province |
CA |
ZIP/Postal code |
94305 |
Country |
USA |
|
|
Platform ID |
GPL4133 |
Series (1) |
GSE14982 |
Reprogramming human fat cells into iPS cells |
|
Relations |
Reanalyzed by |
GSM501454 |