|
| Status |
Public on Dec 10, 2004 |
| Title |
RA_D |
| Sample type |
RNA |
| |
|
| Source name |
rheumatoid arthritis
|
| Organism |
Homo sapiens |
| Characteristics |
sample 1:
age: 41
gender: male
sample 2
age: 66
gender: female
|
| Biomaterial provider |
HELIOS Hospital, Orthopedic Department, Hobrechtsfelder Chaussee 96, D-13125 Berlin
|
| Treatment protocol |
sample 1:
non-steroidal anti-inflammatory drugs (NSAID), disease modifying antirheumatic drugs (DMARD)
sample 2:
disease modifying antirheumatic drugs (DMARD), steroids
|
| Growth protocol |
synovectomy
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated using TRIzol Reagent (Invitrogen) followed by column based clean up (RNeasy, Qiagen).
|
| Label |
alpha 33P-dCTP
|
| Label protocol |
20 microgram of tota RNA was primed with oligodT(12-18) and reverse transcribed with AMV (Roche) in the presence of alpha 33P-dCTP (Amersham Pharmacia Biotech).
|
| |
|
| Hybridization protocol |
The radioactive cDNA was purified using Sephadex columns, denatured and added to denatured salmon sperm and human placenta DNA as blocking reagents. To aid grid positioning during image analysis radioactively labelled 1 ng kanamycin cDNA was added to the complex probe. This mixture was added to 10 ml hybridisation buffer (1M NaCl, 1% SDS, 0.1xSSC) and hybridised overnight at 65°C and then washed three times for 20 min at 65°C (0.1% SDS, 0.1X SSC).
|
| Scan protocol |
The filters were exposed for approx. 6 hours on Fuji BAS screens and signal intensities were read out using a Fuji Bas-1800 reader (Fuji Photo Film).
|
| Description |
pooled synovial tissues from rheumatoid arthritis patients
|
| Data processing |
Signal intensities for every cDNA clone and background spots were determined using the Visual Grid software (GPC-Biotech, Munich). Raw intensities were background corrected by subtracting the local background (estimated block-wise) and normalised.
|
| |
|
| Submission date |
Dec 09, 2004 |
| Last update date |
Feb 21, 2006 |
| Contact name |
Ute Ungethuem |
| E-mail(s) |
[email protected]
|
| Phone |
+49 30 450576272
|
| Organization name |
Charité University Hospital
|
| Department |
Core Center for Functional Genome Research
|
| Street address |
|
| City |
Berlin |
| ZIP/Postal code |
10117 |
| Country |
Germany |
| |
|
| Platform ID |
GPL1740 |
| Series (1) |
|
