|
| Status |
Public on Jun 06, 2007 |
| Title |
6416_Lung developed PGD |
| Sample type |
RNA |
| |
|
| Source name |
lung biopsies from the anterior right middle lobe or lingula immediately prior to cold-flushing.
|
| Organism |
Homo sapiens |
| Characteristics |
P/F ratio at T0:180
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Single isolates of donor lung samples were homogenised in the presence of RNAzolB and finally dissolved in RNase-free H2O. 25 micro-grams of total RNA was treated with DNase using the Qiagen RNase-free DNase kit and samples were further purified using RNeasy spin columns (Qiagen, Valencia, CA). Total RNA treated with DNase was dissolved in RNase-free H2O to a final concentration of 0.2 g/l. RNA quality was assessed by 1% agarose gel electrophoresis in the presence of ethidium bromide. Samples that did not reveal intact and approximately equal 18S and 28S ribosomal bands were excluded from further study.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 25 micrograms total RNA.
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 micrograms of cRNA were hybridized for 16 hr at 45C on GeneChip Human Genome Array (HGU133Av2). GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
Affymetrix GeneArray scanner 3000. Image analysis was performed with the Affymetrix GeneChip software.
|
| Description |
Samples were immediately snap-frozen in liquid nitrogen and then stored in a -70° Celsius freezer until used for analysis. An area of lung tissue approximately 1 x 1 cm was isolated and excised using 2 staple lines from a 30 mm EndoGIA stapler (US Surgical, Norwalk, CT).
|
| Data processing |
The data were normalised using gcRMA. Log2-transformed data.
|
| |
|
| Submission date |
Jun 06, 2007 |
| Last update date |
Jun 06, 2007 |
| Contact name |
Monika Ray |
| E-mail(s) |
mray@cse.wustl.edu
|
| Phone |
3149358788
|
| Fax |
3149356160
|
| Organization name |
Washington University in St. Louis
|
| Street address |
1 Brookings Deive P O Box 1045
|
| City |
St. Louis |
| State/province |
MO |
| ZIP/Postal code |
63112 |
| Country |
USA |
| |
|
| Platform ID |
GPL5356 |
| Series (1) |
| GSE8021 |
Expression data from human donor lung biopsies |
|
