|
| Status |
Public on Jan 01, 2021 |
| Title |
Cells at timepoint Day 7, repeat 2 (29576) |
| Sample type |
RNA |
| |
|
| Source name |
cells treated with osteogenic induction medium at day 7
|
| Organism |
Homo sapiens |
| Characteristics |
repeat id: MSC139
timepoint: D7
|
| Treatment protocol |
MSCs were cultured in MSC growth medium (described above) and used as controls. The treatment (differentiation) protocol involved culturing MSCs in the presence of osteogenic induction (differentiation) medium containing aMEM base medium, 10% fetal bovine serum, 2mM L-glutamine, 100U/ml penicillin, 100µg/ml streptomycin, 10mM β-glycerol phosphate, 50µM ascorbic acid and 10µM dexamethasone.
|
| Growth protocol |
MSCs were cultured and expanded in growth medium containing DMEM (low glucose), 10% fetal bovine serum, 2mM L-glutamine, 100U/ml penicillin, 100µg/ml streptomycin and 1ng/ml basic FGF.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated from whole cell extracts using the Norgen Total RNA purification kit following the manufacturer's protocol.
|
| Label |
biotin
|
| Label protocol |
500 ng of total RNA from whole cell were first subjected to poly-A tailing with Poly A Polymerase and substrate ATP. Then the FlashTag Ligation Mix and ligase were added to carry out the ligation reaction so that the Biotin-labeled, dendritic structutre of 3DNAs were ligated onto miRNA molecules to archieve the miRNA labeling and the amplification of detection signals.
|
| |
|
| Hybridization protocol |
The standard Affymetrix chip hybridization protocol was followed to hybridize the labeled miRNAs onto Affymtrix miRNA array 4.0 in the hybridization oven at 48 C and 60 rpm for 18 hours.
|
| Scan protocol |
After washing and staining, the chips were scanned using the Affymetrix 7G scanner with standard settings to capture the signal intensities for the miRNAs.
|
| Description |
Cells at timepoint Day 7, repeat 2
cellular miRNA expression data
|
| Data processing |
The raw signal data were loaded into Affymetrix Expression Console (v1.4) and subjected to RMA background correction, median summarization of multiple probes, and quantile normalization. This was done only within the probes for Human miRNAs on chip.
|
| |
|
| Submission date |
Apr 03, 2020 |
| Last update date |
Jan 03, 2021 |
| Contact name |
Jinsheng Yu |
| E-mail(s) |
jyu@wustl.edu
|
| Organization name |
Washington University School of Medicine
|
| Department |
Genetics
|
| Lab |
GTAC Lab
|
| Street address |
660 S. Euclid Ave.
|
| City |
St. Louis |
| State/province |
MO |
| ZIP/Postal code |
63110 |
| Country |
USA |
| |
|
| Platform ID |
GPL21572 |
| Series (1) |
| GSE148049 |
Expression profiling of microRNAs during osteogenic differentiation of human MSCs |
|
