|
| Status |
Public on Oct 10, 2007 |
| Title |
14 weeks after first infliximab injection (14-14w-0) |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
14 weeks after first infliximab injection
|
| Organism |
Homo sapiens |
| Characteristics |
Gender: female, Age: 60 years, ACR score at 14 weeks after first infliximab injection: 0%
|
| Extracted molecule |
total RNA |
| Extraction protocol |
The blood (2.5 ml x 2 tubes) was drawn using PAXgene tubes (QIAGEN, Hilden, Germany). The total RNA extraction and purification were carried out according to the manufacturer’s protocol.
|
| Label |
Cy5
|
| Label protocol |
Then total RNA (1 mg) from patients was transcribed and amplified to produce amplified sample RNA (aRNA) using the MessageAmp aRNA kit according to the manufacturer’s instructions. Next, an external control RNA mixture (LD, GP, and RL) was added to both the sample and reference aRNA (9 mg each). These mixed sample and reference aRNA were labeled using SuperScript II kit (Invitrogen, Carlsbad, CA) with random hexamer (TaKaRa, Kyoto, Japan) with Cy3-dUTP and Cy5-dUTP (PerkinElmer, Boston, MA), respectively.
|
| |
|
| Channel 2 |
| Source name |
Reference RNA
|
| Organism |
Homo sapiens |
| Characteristics |
healthy volunteers
|
| Extracted molecule |
total RNA |
| Extraction protocol |
The blood (2.5 ml x 2 tubes) was drawn using PAXgene tubes (QIAGEN, Hilden, Germany). The total RNA extraction and purification were carried out according to the manufacturer’s protocol.
|
| Label |
Cy3
|
| Label protocol |
Then total RNA (1 mg) from patients was transcribed and amplified to produce amplified sample RNA (aRNA) using the MessageAmp aRNA kit according to the manufacturer’s instructions. Next, an external control RNA mixture (LD, GP, and RL) was added to both the sample and reference aRNA (9 mg each). These mixed sample and reference aRNA were labeled using SuperScript II kit (Invitrogen, Carlsbad, CA) with random hexamer (TaKaRa, Kyoto, Japan) with Cy3-dUTP and Cy5-dUTP (PerkinElmer, Boston, MA), respectively.
|
| |
|
| |
| Hybridization protocol |
Competitive hybridization of Cy3-labeled reference and Cy5-labeled sample cDNA on the microarray was carried out according to Brown and Yanofsky[10] with chamber systems (Agilent Technologies, Palo Alto, CA).
|
| Scan protocol |
Slides were scanned five times with five different power ranges using ScanArray 5000 (PerkinElmer, Boston, MA).
|
| Description |
no additional information
|
| Data processing |
The data was converted from tif image data to signal using ImaGene software (BioDiscovery, El Segundo, CA) for further statistical analysis. Five file data of each four spot data corresponding each gene were merged to establish the single representative data for each gene (Patent pending: PCT/JP03/06677).
|
| |
|
| Submission date |
Jul 02, 2007 |
| Last update date |
Aug 14, 2011 |
| Contact name |
Tsutomu Takeuchi |
| Organization name |
Saitama Medical University
|
| Department |
Department of Internal Medicine
|
| Lab |
Division of Rheumatology/Clinical Immunology
|
| Street address |
1981 Tsujido-machi, Kamoda
|
| City |
Kawagoe-shi, Saitama |
| ZIP/Postal code |
350-8550 |
| Country |
Japan |
| |
|
| Platform ID |
GPL5460 |
| Series (1) |
| GSE8350 |
mRNA expression profile in peripheral blood cells of RA patients following treatment with an anti-TNFa mAb, infliximab |
|
