|
| Status |
Public on Dec 26, 2018 |
| Title |
hADSC06 P(25) |
| Sample type |
RNA |
| |
|
| Source name |
hADSC (adult)
|
| Organism |
Homo sapiens |
| Characteristics |
condition: Adult
passage: Long
|
| Treatment protocol |
No specific treatment; compared samples were cultured for long and short passages.
|
| Growth protocol |
Cells were cultured in DMEM supplemented with 10% FBS, 2 mM glutamine, penicillin (100 U/ml)/streptomycin (1000 U/ml) (37ºC, 5% CO2); 3% Oxygen
|
| Extracted molecule |
total RNA |
| Extraction protocol |
mirVana miRNA Isolation Kit (Applied Biosystems)
|
| Label |
Cy3
|
| Label protocol |
miRNA Microarray System Protocol (Agilent Technologies) was used to label RNA; 100 ng of total RNA were dephosphorylated and Cyanine 3-pCp molecule was ligated to the 3« end of each RNA molecule by using T4 RNA ligase.
|
| |
|
| Hybridization protocol |
100 ng of Cy3 labelled RNA were hybridized for 20 hours at 55¼C in a hybridization oven (G2545A, Agilent) set to 15 rpm in a final concentration of 1X GE Blocking Agent and 1X Hi-RPM Hybridization Buffer, according to manufacturer's instructions (miRNA Microarray System Protocol, Agilent Technologies)
|
| Scan protocol |
Arrays were scanned at 5μm resolution on an Agilent DNA Microarray Scanner (G2565BA, Agilent Technologies) using the default settings for miRNA Microarray v1.0 (miRNA Microarray System Protocol, Agilent Technologies).
|
| Description |
Expression of miRNAs from hADSC cell culture
|
| Data processing |
Images provided by the scanner were analyzed using Agilent´s software Feature Extraction version 9.5.3.1.
Processed Cy3 signal intensity data was normalized based on the VSN-invariant method (Huber 2002, Pradervand 2009)
|
| |
|
| Submission date |
Oct 18, 2018 |
| Last update date |
Dec 26, 2018 |
| Contact name |
Fatima Sanchez-Cabo |
| E-mail(s) |
fscabo@cnic.es
|
| Phone |
+34 91 4531200
|
| Organization name |
CNIC
|
| Street address |
Melchor Fernandez Almagro
|
| City |
Madrid |
| ZIP/Postal code |
28029 |
| Country |
Spain |
| |
|
| Platform ID |
GPL7731 |
| Series (1) |
| GSE121481 |
microRNA profiles of hADSC during replicative senescence |
|
