|
| Status |
Public on Jul 30, 2014 |
| Title |
Blood-Volunteer5 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
blood sample at the beginning of the study (time 0)
|
| Organism |
Homo sapiens |
| Characteristics |
biological sources: blood
mof supplementation: none
gender: male
age: 30
cigarettes/day: 16.5
years of smoking: 15
height (m): 1.88
weight (kg): 93.2
bmi (kg/m2): 26.4
sys bp (mm hg): 114
dia bp (mm hg): 62
heart rate (beats/min): 53
|
| Treatment protocol |
At baseline and after 8 weeks of flavanol supplementation freshly collected blood was added to RNALater® (Ambion, Austin, TX, USA) and stored at -80°C.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated from blood by means of the RiboPure-Blood kit (Ambion, Austin, TX, USA) in accordance with the manufacturer’s instructions.
|
| Label |
Cy3
|
| Label protocol |
50 ng of total RNA extracted were amplified and fluorescently labelled to produce Cy5 or Cy3 cRNA using the Low Input Quick Amp Labeling two colors Kit (Agilent, USA) in the presence of spike-in two colors control.
|
| |
|
| Channel 2 |
| Source name |
blood samples after 8 weeks of MOF supplementation
|
| Organism |
Homo sapiens |
| Characteristics |
biological sources: blood
mof supplementation: 8 weeks
gender: male
age: 30
cigarettes/day: 16.5
years of smoking: 15
height (m): 1.88
weight (kg): 93.2
bmi (kg/m2): 26.4
sys bp (mm hg): 114
dia bp (mm hg): 62
heart rate (beats/min): 53
|
| Treatment protocol |
At baseline and after 8 weeks of flavanol supplementation freshly collected blood was added to RNALater® (Ambion, Austin, TX, USA) and stored at -80°C.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated from blood by means of the RiboPure-Blood kit (Ambion, Austin, TX, USA) in accordance with the manufacturer’s instructions.
|
| Label |
Cy5
|
| Label protocol |
50 ng of total RNA extracted were amplified and fluorescently labelled to produce Cy5 or Cy3 cRNA using the Low Input Quick Amp Labeling two colors Kit (Agilent, USA) in the presence of spike-in two colors control.
|
| |
|
| |
| Hybridization protocol |
After purification (Qiagen RNeasy Mini Kit), 825 ng of Cy3 and Cy5 labeled cRNA were hybridized onto G4845A Human GE 4x44K v2 microarray (Agilent, USA) with hybridization buffer (Agilent Gene Expression Hybridization Kit) according to the manufacturer’s instructions. After 17h of hybridization, slides were washed with the Gene Expression Wash Pack.
|
| Scan protocol |
Microarrays were scanned with Agilent G2505 scanner.
|
| Data processing |
Data were extracted with Feature Extraction 10.7 software (Agilent, USA) using Lowess normalization (no background subtraction).
|
| |
|
| Submission date |
Jan 23, 2014 |
| Last update date |
Jul 30, 2014 |
| Contact name |
Celine BOBY |
| Organization name |
INRA
|
| Street address |
Centre INRA Auvergne-Rhône-Alpes
|
| City |
Saint Genes Champanelle |
| ZIP/Postal code |
63122 |
| Country |
France |
| |
|
| Platform ID |
GPL10332 |
| Series (2) |
| GSE54325 |
Impact of monomeric and oligomeric flavanols (MOF) consumption on the gene expression profile of leukocytes (human) |
| GSE54690 |
Transcriptome-Methylome study of blood samples from flavanol diet intervention Paper title: Dietary flavanols modulate the transcription of genes associated with cardiovascular pathology and induces complex changes of their DNA methylation state. |
|
