|
| Status |
Public on May 18, 2012 |
| Title |
Monocyte_obese_rep5 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Human, CD14+ monocytes, obese
|
| Organism |
Homo sapiens |
| Characteristics |
gender: female
cell type: CD14+ monocytes
disease state: obese
gender: female
cell type: CD14+ monocytes
disease state: lean-obese
|
| Growth protocol |
Blood samples were collected and after removal of the plasma fraction, peripheral blood mononuclear cells (PBMCs) were isolated using gradient separation on Histopaque-1077. CD14+ monocytes were isolated from the PBMC fraction using CD14 immunomagnetic beads (Miltenyi).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted with Trizol reagent, followed by clean-up and DNase I treatment with Qiagen miRNeasy mini kit in accordance with the prescribed protocol provided with the kit. Quality control was performed with Agilent Bioanalyzer.
|
| Label |
Hy3
|
| Label protocol |
600 ng total RNA from sample and reference pool was labelled with Hy3 and Hy5 fluorescent label, respectively, using the miRCURY LNA Array power labelling kit (Exiqon).
|
| |
|
| Channel 2 |
| Source name |
Human, CD14+ monocytes, lean-obese reference pool
|
| Organism |
Homo sapiens |
| Characteristics |
gender: female
cell type: CD14+ monocytes
disease state: obese
gender: female
cell type: CD14+ monocytes
disease state: lean-obese
|
| Growth protocol |
Blood samples were collected and after removal of the plasma fraction, peripheral blood mononuclear cells (PBMCs) were isolated using gradient separation on Histopaque-1077. CD14+ monocytes were isolated from the PBMC fraction using CD14 immunomagnetic beads (Miltenyi).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted with Trizol reagent, followed by clean-up and DNase I treatment with Qiagen miRNeasy mini kit in accordance with the prescribed protocol provided with the kit. Quality control was performed with Agilent Bioanalyzer.
|
| Label |
Hy5
|
| Label protocol |
600 ng total RNA from sample and reference pool was labelled with Hy3 and Hy5 fluorescent label, respectively, using the miRCURY LNA Array power labelling kit (Exiqon).
|
| |
|
| |
| Hybridization protocol |
The hybridization was performed according to the miRCURY LNA array manual using a Tecan HS4800 hybridization station.
|
| Scan protocol |
The microarray slides were scanned using the Agilent G2565BA Microarray Scanner System and the image analysis was carried out using the ImaGene 8.0 software (BioDiscovery).
|
| Data processing |
The quantified signals were background corrected (Normexp with offset value 10) and normalized using the global LOWESS (LOcally WEighted Scatterplot Smoothing) regression algorithm.
|
| |
|
| Submission date |
Dec 07, 2011 |
| Last update date |
May 18, 2012 |
| Contact name |
Paul Holvoet |
| E-mail(s) |
[email protected]
|
| URL |
http://www.kuleuven.be/amu
|
| Organization name |
K.U.Leuven
|
| Department |
Department of Cardiovascular Diseases
|
| Lab |
Atherosclerosis and Metabolism Unit
|
| Street address |
Herestraat 49, PB 705
|
| City |
Leuven |
| ZIP/Postal code |
3000 |
| Country |
Belgium |
| |
|
| Platform ID |
GPL11432 |
| Series (1) |
| GSE34223 |
microRNA expression analysis of circulating monocytes in obese patients |
|
