|
| Status |
Public on Aug 22, 2019 |
| Title |
H9_substate 2_exp3 |
| Sample type |
RNA |
| |
|
| Source name |
H9, passage 65, substate 2, miRNA from the cells, experiment 3
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: human embryonic stem cell
cell line: H9
|
| Growth protocol |
H9 cells were maintained on Matrigel (BD Biosciences) in TeSR-E8 (STEMCELL Technologies). The cultures were incubated at 37 ºC in a humidified incubator with 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
miRNA was prepared by using miRNeasy Mini Kit (QIAGEN, 217004) following the manufacturer's recommendations. The miRNA was quantified by a Agilent bioanalyzer, using the RNA 6000 Pico Kit (Agilent, 5067-1513).
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 100 ng of miRNA using miRNA Complete Labeling and Hyb Kit (Agilent, 5190-0456) according to the manufacturer's instructions, followed by MicroBioSpin6 column purification (Bio Rad, 732-62221).
|
| |
|
| Hybridization protocol |
The Cy3-labelled cRNA was dried up with vacuum concentrator, and then was dissolved in hybridization buffer (Agilent, 5190-0456) according to the manufacturer's instructions. The sample was hybridized to SurePrint G3 miRNA Microarray Kit, 8 x 60K (Agilent, G4872A) for 20 hours at 55°C in a rotating Agilent hybridization oven. After hybridization, microarrays were washed 5 minute at room temperature with GE Wash Buffer 1 (Agilent) and 5 minute with 37°C GE Wash buffer 2 (Agilent).
|
| Scan protocol |
Slides were scanned immediately after washing on the Agilent DNA Microarray Scanner (G2505C) using one color scan setting for 8x60k array slides.
|
| Description |
miRNA expression in human embryonic stem cell
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software GX14.9.1 (Agilent) using default parameters.
|
| |
|
| Submission date |
Dec 06, 2018 |
| Last update date |
Aug 22, 2019 |
| Contact name |
Yuzuru Ito |
| Organization name |
National Institute of Advanced Industrial Science and Technology (AIST)
|
| Department |
Biotechnology Research Institute for Drug Discovery
|
| Street address |
Central 6, Higashi 1-1-1
|
| City |
Tsukuba |
| State/province |
Ibaraki |
| ZIP/Postal code |
3058566 |
| Country |
Japan |
| |
|
| Platform ID |
GPL25134 |
| Series (1) |
| GSE123466 |
Expression profiles of miRNA derived from cells for ectoderm-biased substate of human pluripotent stem cells. |
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