|
Status |
Public on Dec 30, 2022 |
Title |
CTR 46 |
Sample type |
RNA |
|
|
Source name |
Plasma extracellular vesicles
|
Organism |
Homo sapiens |
Characteristics |
tissue: Plasma extracellular vesicles disease state: healthy control molecule subtype: microRNA
|
Extracted molecule |
total RNA |
Extraction protocol |
Extracellular vesicles (EV) isolation and RNA extraction from plasma and synovial fluid was performed with the exoRNeasy Serum/Plasma Midi kit (Qiagen Italia, Milano, Italy).
|
Label |
Applied Biosystems Human TaqMan Low Density Array (TLDA, v2.0) (Card A)
|
Label protocol |
EV-miR were analyzed by the TaqMan Array Card Technology. EV-miR were reverse transcribed with the TaqMan microRNA Reverse Transcription Kit, using the Megaplex RT primers Human Pool A (Thermo Fisher Scientific, Monza, MB, Italy). Pre-amplification of cDNA was performed with TaqMan PreAmp Master Mix and MegaplexTM Pre-Amp primers Human Pool A. The pre-amplification product was diluted according to the manufacturer’s instructions and used to perform microRNA profiling on the ViiA 7 Real-Time PCR System.
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|
|
Hybridization protocol |
n/a
|
Scan protocol |
n/a
|
Description |
Healthy controls
|
Data processing |
Data processing, categorization, normalization, filtering, imputation and differential expression was carried out by PIPE-T galaxy tool. Ct values falling within the range 14-32 were categorized as reliable values as recommended by the manufacturer guidelines. Global mean normalization was used to reduce any technical variability introduced in the data by the RT-qPCR experiments. Only Exo-miR with at most 20% of missing values were retained for the analysis to reduce the bias introduced by imputation. Mestdagh method was employed to assign a numeric expression value to missing values. The fold change and t test method was used to identify significant differentially expressed microRNA. non-normalized data file reports raw Ct values. Ct value of 40 indicates a missing value. Normalized data file reports delta Ct values (Ct – sample mean expression ). NA value indicates a missing value.
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|
|
Submission date |
Dec 20, 2021 |
Last update date |
Dec 30, 2022 |
Contact name |
Federica Raggi |
E-mail(s) |
federicaraggi@gaslini.org
|
Organization name |
IRCCS Istituto Giannina Gaslini
|
Lab |
Molecular biology
|
Street address |
via genrolamo gaslini, 5
|
City |
Genova |
ZIP/Postal code |
16147 |
Country |
Italy |
|
|
Platform ID |
GPL19079 |
Series (1) |
GSE191326 |
Extracellular vesicle microRNA profiling in synovial fluids and plasma from Oligoarticular Juvenile Idiopathic Arthritis patients: methodological challenges and identification of putative disease biomarkers |
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