|
| Status |
Public on Jul 26, 2022 |
| Title |
Healthy PBMC child h |
| Sample type |
RNA |
| |
|
| Source name |
Human mononuclear cells from healthy donors isolated from peripheral blood
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: mononuclear cells from peripheral blood
disease state: healthy donor
age: 8 years old
developmental stage: children
|
| Treatment protocol |
not applicable
|
| Growth protocol |
Human mononuclear cells from patients with JIA were isolated onto a Ficoll (PAN Biotech, Germany) gradient either from peripheral blood (PB) or synovial fluid (SF). Erythrocytes were lysed and cells were washed twice. Peripheral blood mononuclear cells (PBMC) were isolated from healthy donors by the same procedure. SF was obtained by incubation with hyaluronidase for 30 min. Following centrifugation at 400 x g for 10 min.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
miRNA extraction kit preAnalytix plus DNase digstion
|
| Label |
biotin
|
| Label protocol |
1.5 µg of total RNA containing low molecular weight RNA was labelled using the FlashTag™ Biotin HSR RNA Labeling Kit according to the manufacturer’s instructions (ThermoFisher Scientific).
|
| |
|
| Hybridization protocol |
Each sample was hybridized to a GeneChip® miRNA 3.0 Array for 18 h at 48°C and 60 rpm (Affymetrix, Santa Clara, CA, USA). Afterwards, the chips were washed and stained on Fluidics Station 450 (Fluidics script FS450-0002).
|
| Scan protocol |
Arrays were scanned on a GeneChip® Scanner 3000 7G (both, Affymetrix, Santa Clara, CA, USA).
|
| Description |
miRNA
|
| Data processing |
Data were analyzed by using expression console software 1.4 (Affymetrix, Santa Clara, CA, USA). The expression values were normalized with robust multi-array average (RMA).miRNA calculations and statistical significances between two groups (paired Student’s t-test) were performed with Transcriptome Analysis Console (TAC) Software (ThermoFisher Scientific). Detected miRNAs whose expression levels were significantly different (FDR-p value < 0.01) and showed a linear-fold change of at least 3.0, were considered as differentially expressed.
|
| |
|
| Submission date |
Jul 27, 2019 |
| Last update date |
Jul 26, 2022 |
| Contact name |
Bernd Denecke |
| E-mail(s) |
bernd.denecke@rwth-aachen.de
|
| Phone |
+49 241 8089918
|
| Organization name |
RWTH Aachen University
|
| Department |
IZKF Aachen
|
| Street address |
Pauwelsstrasse 30
|
| City |
Aachen |
| State/province |
NRW |
| ZIP/Postal code |
52074 |
| Country |
Germany |
| |
|
| Platform ID |
GPL16384 |
| Series (1) |
| GSE134984 |
miR-23a regulates T cellular Redox metabolism in juvenile idiopathic arthritis |
|
