|
| Status |
Public on Aug 23, 2008 |
| Title |
Endothelial cells from left iliac artery, rep 2 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
left iliac endothelial cells
|
| Organism |
Sus scrofa |
| Characteristics |
Animal 2
|
| Growth protocol |
Reference endothelial cells were grown in M199 supplemented with antibiotic/mycotic, FBS, and porcine serum. Passage 3 cells were used.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Qiagen RNeasy Micro kit following manufacturer's protocol
|
| Label |
Cy5
|
| Label protocol |
Messenger RNA was amplified using a MessageAmp II aRNA amplification kit (Ambion). two rounds of amplification were performed, and aminoallyl-UTP was used in the final round to provide reactive groups for dye incorporation. The RNA samples were labeled with Cy5 dyes, and reference RNA sample was labeled with Cy3 dyes.
|
| |
|
| Channel 2 |
| Source name |
reference RNA
|
| Organism |
Sus scrofa |
| Characteristics |
cultured porcine aortic endothelial cells (passage 3)
|
| Growth protocol |
Reference endothelial cells were grown in M199 supplemented with antibiotic/mycotic, FBS, and porcine serum. Passage 3 cells were used.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Qiagen RNeasy Micro kit following manufacturer's protocol
|
| Label |
Cy3
|
| Label protocol |
Messenger RNA was amplified using a MessageAmp II aRNA amplification kit (Ambion). two rounds of amplification were performed, and aminoallyl-UTP was used in the final round to provide reactive groups for dye incorporation. The RNA samples were labeled with Cy5 dyes, and reference RNA sample was labeled with Cy3 dyes.
|
| |
|
| |
| Hybridization protocol |
• Make hybridization buffer fresh each time. • Add 38 ul hybridization buffer to 12 ul fragmented Cy Dye labeled aa-aRNA. • Add 1ul each poly dA and appropriate Cot-1 DNA. • Heat at 95ºC for 2 minutes and put on ice to cool down. • Mix well and load sample to the array slide with lifter cover slip. • Hybridize overnight in 42ºC water bath.
|
| Scan protocol |
Scanned by GenePix 4000B and quantified using GenePix software
|
| Description |
Endothelial cells were freshly harvest from right coronary, left and right iliac arteries from four pigs.
|
| Data processing |
LOWESS normalized, inlcude genes with signal in at least 8 of the 12 arrays, Agilent GeneSpring GX was used.
|
| |
|
| Submission date |
Mar 25, 2008 |
| Last update date |
Aug 23, 2008 |
| Contact name |
Ji Zhang |
| E-mail(s) |
ji.z@duke.edu
|
| Phone |
919-660-5125
|
| Fax |
919-684-4488
|
| Organization name |
Duke University
|
| Department |
Department of Biomedical Engineering
|
| Lab |
Cardiovascular simulations
|
| Street address |
Room 136 Hudson Hall
|
| City |
Durham |
| State/province |
NC |
| ZIP/Postal code |
27708 |
| Country |
USA |
| |
|
| Platform ID |
GPL3461 |
| Series (1) |
| GSE10938 |
In vivo endothelial transcriptional profiles of porcine coronary and iliac arteries |
|
