|
| Status |
Public on Nov 01, 2011 |
| Title |
Aging muscular sample 2 Replicate 2 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
AGE
|
| Organism |
Homo sapiens |
| Characteristics |
gender: Female
age (years): 65
muscle: Paravertebral
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Normal and patient total RNA was purified using Trizol Reagent (Life technologies). Poly (A)+ RNA was purified from total RNA using oligo-dt linked oligotex resin (QIAGEN).
|
| Label |
Cy5
|
| Label protocol |
Cy3- and Cy5-labeled cDNA was prepared using the CyScribe cDNA post labeling Kit (Amersham Pharmacia Biotech). Samples from DMD and AGING patients were each labeled individually with Cy5. Reference patients were pooled and then labeled with Cy3.
|
| |
|
| Channel 2 |
| Source name |
CONTROL
|
| Organism |
Homo sapiens |
| Characteristics |
description: REFERENCE: pool of total RNA from 7 younger CONTROL muscle biopsies
gender: 5 Females / 2 Males
age (years): 14-30
muscle: Paravertebral
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Normal and patient total RNA was purified using Trizol Reagent (Life technologies). Poly (A)+ RNA was purified from total RNA using oligo-dt linked oligotex resin (QIAGEN).
|
| Label |
Cy3
|
| Label protocol |
Cy3- and Cy5-labeled cDNA was prepared using the CyScribe cDNA post labeling Kit (Amersham Pharmacia Biotech). Samples from DMD and AGING patients were each labeled individually with Cy5. Reference patients were pooled and then labeled with Cy3.
|
| |
|
| |
| Hybridization protocol |
Microarrays were prehybridized for 1 hour at 42° C (in 10% BSA, 20XSSC, 10% SDS). 0.5 µg of labeled sample cDNA was mixed with 0.5 µg of labeled reference cDNA in 40 µl of hybridization mix (50X denhardt, 20XSSC, 10 µg/µl PolyA RNA, 10 µg/µl Yeast tRNA, 10% SDS, 50% formamide) and hybridized to the microarrays. Microarray hybridization was carried out overnight at 42° C in a humidified chamber. Finally, slides were washed and dried by centrifugation.
|
| Scan protocol |
Array scanning was carried out using a GSI lumonics LITE dual confocal laser scanner with ScanArray Microarray Analysis Software. Raw scanner images were analyzed with Genepix Software (AXON LABORATORY).
|
| Description |
AGE, Biological replicate 2 of 4, Technical replicate 2 of 2.
|
| Data processing |
Data were normalized (LOWESS) and analyzed (data filtering) using the Micro-Array Data Suite of Computed Analysis (MADSCAN)
|
| |
|
| Submission date |
Oct 10, 2011 |
| Last update date |
Nov 01, 2011 |
| Contact name |
Daniel BARON |
| E-mail(s) |
[email protected]
|
| Organization name |
Institut national de la santé et de la recherche médicale-INSERM
|
| Department |
UMR1064 - Centre de Recherche en Transplantation & Immunologie
|
| Lab |
Tolerance et regulation lymphocytaire
|
| Street address |
30 Bd Jean Monnet
|
| City |
Nantes |
| State/province |
Loire-Atlantique (44) |
| ZIP/Postal code |
44093 |
| Country |
France |
| |
|
| Platform ID |
GPL14713 |
| Series (1) |
|
