|
| Status |
Public on Mar 14, 2017 |
| Title |
reMAIT_day_10_Middle |
| Sample type |
RNA |
| |
|
| Source name |
reMAIT, day 10, Middle
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: cord blood
timecourse: 7-10 day
|
| Biomaterial provider |
Hokkaido Cord Blood Bank
|
| Treatment protocol |
Cells were harvested with TRIzol (Sigma, Aldrich).
|
| Growth protocol |
MAIT-derived iPSCs were cultured on OP9, and CD34+ CD43+ cells were isolated. These CD34+ CD43+ precursor cells were furthered cultured on OP9/DL1 for re-differentiation into MAITs.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was prepared using the RNAeasy column purification (QIAGEN, Valencia, CA) following the manufacturer's recommendations. RNA was quantified using a NanoDrop-1000 spectrophotometer and quality was monitored with the Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA).
|
| Label |
Cy3-pCp
|
| Label protocol |
Sample labeling was performed as detailed in the “miRNA Microarray System with miRNA Complete Labeling and Hyb Kit” protocol (version 2.4, part number G4170-90011). Briefly, 90 ng total RNA of sample hES d0 Pool and 100 ng total RNA of all other samples was used for the labeling step using the miRNA Complete Labeling and Hyb Kit (Agilent Technologies).
|
| |
|
| Hybridization protocol |
The hybridization procedure was performed according to the “miRNA Complete Labeling and Hyb Kit” protocol using the miRNA Complete Labeling and Hyb Kit (Agilent Technologies). Briefly, Cy3-labeled RNA in hybridization buffer was hybridized overnight (20 hours, 55 °C) to Agilent Human microRNA Microarrays 8x60K v19 (Design ID 046064) using Agilent’s recommended hybridization chamber and oven. Following hybridization, the microarrays were washed once with the Agilent Gene Expression Wash Buffer 1 for 5 min at room temperature followed by a second wash with preheated Agilent Gene Expression Wash Buffer 2 (37 °C) for 5 min.
|
| Scan protocol |
Fluorescence signals of the hybridized Agilent Microarrays were detected using Agilent’s Microarray Scanner System (G2505C, Agilent Technologies, Palo Alto, USA).
|
| Description |
Micro RNA expression from reMAIT at 7-10 days after the beginning of induction
|
| Data processing |
The Agilent Feature Extraction Software (FES 10.7.3.1 ) was used to read out and process the microarray image files.
|
| |
|
| Submission date |
Oct 19, 2016 |
| Last update date |
Mar 14, 2017 |
| Contact name |
Toutai MITUYAMA |
| Organization name |
National Institute of Advanced Industrial Science and Technology (AIST)
|
| Department |
Artificial Intelligence Research Team
|
| Lab |
Computational Omics Research Team
|
| Street address |
2-4-7 Aomi
|
| City |
Koto-ku |
| State/province |
Tokyo |
| ZIP/Postal code |
135-0064 |
| Country |
Japan |
| |
|
| Platform ID |
GPL19730 |
| Series (2) |
| GSE88936 |
Time series micro RNA analysis of immature T-cells and reMAIT cells by microarrays |
| GSE88938 |
Time series transcriptome and DNA methylome analysis of immature T-cells and reMAIT cells |
|
