|
| Status |
Public on Feb 04, 2013 |
| Title |
104_after PCB_rep2 |
| Sample type |
RNA |
| |
|
| Source name |
after 8 wks of placebo (PCB) diet
|
| Organism |
Homo sapiens |
| Characteristics |
diet group: PCB
tissue: skeletal muscle
sample_id: 104B
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was prepared using the mirVanaTM miRNA Isolation Kit (Applied Biosystems) following the manufacturer's recommendations. RNA was quantified using a NanoDrop-1000 spectrophotometer and quality was monitored with the Agilent 2100 Bioanalyzer (Agilent Technologies).
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 0.2 µg RNA using the One-Color Low Input Quick Amp labeling (Agilent) according to the manufacturer's instructions, followed by purification with Absolutely RNA Nanoprep kit (Stratagen). Dye incorporation and cRNA yield were checked with the NanoDrop ND-1000 Spectrophotometer.
|
| |
|
| Hybridization protocol |
1.65 ug of Cy3-labelled cRNA was fragmented at 60°C for 30 minutes in a reaction volume of 55µl containing 2x Agilent fragmentation buffer and 10x Agilent blocking agent following the manufacturers instructions. On completion of the fragmentation reaction, 55µl of 2x Agilent hybridization buffer was added to the fragmentation mixture and hybridized to Agilent Whole Human Genome Oligo Microarrays (G4845A) for 17 hours at 65°C in a rotating Agilent hybridization oven. After hybridization, microarrays were washed 1 minute at room temperature with GE Wash Buffer 1 (Agilent) and 1 minute with 37°C GE Wash buffer 2 (Agilent), then dried immediately by brief centrifugation.
|
| Scan protocol |
Slides were scanned immediately after washing on the Agilent DNA Microarray Scanner using one color scan setting for 4x44k array slides.
|
| Description |
Gene Expression after 8 weeks of PCB diet
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 10.5 (Agilent) using default parameters to obtain background subtracted median normalized Signal intensities.
|
| |
|
| Submission date |
Feb 13, 2012 |
| Last update date |
Feb 04, 2013 |
| Contact name |
emmanuelle meugnier |
| E-mail(s) |
meugnier@univ-lyon1.fr
|
| Organization name |
INSERMU1060/INRA1397
|
| Lab |
CarMeN
|
| Street address |
Cens Eli Bat 2 D- Hopital Lyon Sud
|
| City |
Pierre Bénite |
| ZIP/Postal code |
69495 |
| Country |
France |
| |
|
| Platform ID |
GPL13497 |
| Series (1) |
| GSE35764 |
Polyphenols protection against fructose overload |
|
