|
| Status |
Public on Feb 10, 2007 |
| Title |
Conjunctiva Sample 3 |
| Sample type |
RNA |
| |
|
| Source name |
Fresh Human Conjunctiva
|
| Organism |
Homo sapiens |
| Characteristics |
This sample comprises of RNA isolated from human conjunctivae pooled from a mixed population of (age, sex and cause of death identified). Tissue was send to us by the National Disease Resource Center (NDRI); http://www.ndriresource.org/
|
| Biomaterial provider |
National Disease Resource Center (NDRI), Philadelphia, USA.
|
| Treatment protocol |
None. Fresh tissue from donor, post mortem
|
| Growth protocol |
N/A Fresh tissue
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Epithelial sheets were manually transferred from the Dispase solution directly into lysis buffer, total RNA was isolated using the RNeasy minikit (Qiagen Inc., Valencia, CA) and was further purified using RNeasy (Qiagen) spin columns. RNA was suspended in RNase-Free water at a concentration of 1.5 ug/ul
|
| Label |
Biotin-labled cRNA
|
| Label protocol |
Biotin labeled cRNA using the ENZO RNA transcript labeling Kit 9 (BioArray High Yeald Labeling Kit, P/N 900182). IVT cRNA cleanup and quantitation, cRNA fragmentation, control targets performed according to Affymetrix protocols. Synthesis of cDNA using T7-(dT)24 Primer (Genset Corp) and Superscript Choice System (Gibco BRL Life technologies).
|
| |
|
| Hybridization protocol |
Hybridization cocktail made according to Affymetrix protocols. Heated to 99C and then heat block for 5 mins at 45oC. Incubate probe array with 1X hybridization buffer (MES, 1M [Na], 20mM EDTA, 0.01% Tween) for 10 mins 45oC. Hybridize for 16 h 45oC using final volume of 200ul. For washing use GeneChip Fluidics Station 450 using fluidics protocol EukGE-WS2v4. Hybridized microarrays were stained with a strepavidin-phycoerythrin (SAPE) reagent.
|
| Scan protocol |
Hybridized microarrays were stained with a strepavidin-phycoerythrin (SAPE) reagent and fluorescence images were captured using an Agilent G2500A laser scanner.
|
| Description |
Standard Affymetrix protocols performed
|
| Data processing |
Microarray Analysis Suite, version 5.0 (MAS 5.0; Affymetrix)
|
| |
|
| Submission date |
Aug 10, 2006 |
| Last update date |
Aug 15, 2006 |
| Contact name |
Jose Mario Wolosin |
| E-mail(s) |
jose.wolosin@mssm.edu
|
| Phone |
212 241 8980
|
| Organization name |
Mount Sinai School of Medicine
|
| Department |
Ophthalmology
|
| Lab |
22-76
|
| Street address |
1 Gustave L. Levy Place
|
| City |
New York |
| State/province |
NY |
| ZIP/Postal code |
10029 |
| Country |
USA |
| |
|
| Platform ID |
GPL96 |
| Series (1) |
| GSE5543 |
Gene Expression in Human Conjunctiva and Cornea |
|
