|
| Status |
Public on Oct 18, 2022 |
| Title |
SubjectA_Pre-1_Exp1 |
| Sample type |
RNA |
| |
|
| Source name |
Whole blood, Pre -1, SubjectA, Experiment1
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: Whole blood
gender: male
condition: pre-confinement
subject: A
|
| Treatment protocol |
Blood samples were collected in tubes at 15:00-16:00 on 1 day before staying at isolation and confinement facility and day 14th during. Whole blood samples were incubated at room temperature for 24 hours after collecting and stored at -80℃ until analysis.
|
| Growth protocol |
N/A
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was extracted using the PAXgene Blood RNA System Kit (Qiagen, Tokyo, Japan) according to the manufacturer’s guidelines.
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled aRNA was prepared from RNA using the Low Input Quick Amp Labeling Kit and the RNA Spike-In Kit (1 color) (Agilent Technologies) according to the manufacturer's instructions, followed by RNeasy column purification (QIAGEN, Valencia, CA). Dye incorporation and cRNA yield were checked with the NanoDrop ND-1000 Spectrophotometer.
|
| |
|
| Hybridization protocol |
1.5 ug of Cy3-labelled cRNA (specific activity >10.0 pmol Cy3/ug cRNA) was fragmented at 60°C for 30 minutes in a reaction volume of 0.1ml containing 1x Agilent fragmentation buffer and 2x Agilent blocking agent following the manufacturer's instructions. On completion of the fragmentation reaction, 0.1 ml of 2x Agilent hybridization buffer was added to the fragmentation mixture and hybridized to Agilent Human Gene Expression 4x44K v2 Microarray Kit (G4845A) for 17 hours at 65°C in a rotating Agilent hybridization oven. After hybridization, microarrays were washed 1 minute at room temperature with GE Wash Buffer 1 (Agilent) and 1 minute with 37°C GE Wash buffer 2 (Agilent), then dried immediately by brief centrifugation.
|
| Scan protocol |
Slides were scanned on the Agilent DNA Microarray Scanner (G2505B).
|
| Description |
A_Pre-1_1
Gene expression in Pre -1 human whole blood.
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software (Agilent) using default parameters to obtain background subtracted and spatially detrended Processed Signal intensities. Features flagged in Feature Extraction as Feature Non-uniform outliers were excluded.
|
| |
|
| Submission date |
Sep 07, 2017 |
| Last update date |
Oct 18, 2022 |
| Contact name |
Masaya Seki |
| E-mail(s) |
seki.masaya@jaxa.jp
|
| Organization name |
Japan Aerospace Exploration agency
|
| Department |
JEM Utilization Center
|
| Street address |
Sengen 2-1-1
|
| City |
Tsukuba |
| State/province |
Ibaraki |
| ZIP/Postal code |
305-8505 |
| Country |
Japan |
| |
|
| Platform ID |
GPL13497 |
| Series (1) |
| GSE103605 |
Effects of 14 days of confinement on blood gene expression profiles in men |
|
