|
| Status |
Public on Aug 31, 2016 |
| Title |
microG_4days_rep3 |
| Sample type |
RNA |
| |
|
| Source name |
microgravity for 4days, replicate 3
|
| Organism |
Caenorhabditis elegans |
| Characteristics |
strain: N2 wild type
Stage: Adult
|
| Treatment protocol |
The worms were washed by M9 buffer. The worms were homoginaized with four rounds of 20-secound beating at 5000rpm in a MicroSmash MS-100 bead beater (Tomy seiko,Tokyo JAPAN).
|
| Growth protocol |
The worms were cultured at 20 degree for 4 days in internatioal space station (ISS). The 2 mL of M9 cholesterol buffer suspended 9,000 of the hatched L1 larvae and the 12 mL of S-basal media suspended E. coli (OD600=5) were separately packed into a monolayer polyethylene bag. The experiment was activated once onboard the ISS by reintroducing worms to bacterial food.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was prepared using ISOGEN (Nippon Gene, Tokyo, Japan) following the manufacturer's recommendations. The residual DNA was eliminated by treating with DNase I (Takara Bio, Shiga, Japan) during an isolation process of total RNA. RNA was quantified using a NanoDrop-1000 spectrophotometer and quality was monitored with the Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA).
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 0.5 ug RNA using the One-Color Low RNA Input Linear Amplification kit (Agilent) according to the manufacturer's instructions, followed by RNAeasy column purification (QIAGEN, Valencia, CA).
|
| |
|
| Hybridization protocol |
The labeled cRNA (1.65 µg/sample) was hybridized to a microarray using the Agilent Gene Expression Hybridization Kit at 65°C for 17 hours.
|
| Scan protocol |
Slides were scanned immediately after washing on the Agilent DNA Microarray Scanner (G2505C) using one color scan setting for 4x44k array slides.
|
| Description |
Gene expression in microG
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 10.7.3.1.
Microarray data were analyzed using Gene Spring software (Agilent Technologies). Normalized data were produced by Normalize to 75 percentile sift protocol’’ and baseline to median of control sample protocol’’. Arrays were filtered on expression (20-100)th percentile in raw data.
|
| |
|
| Submission date |
Aug 06, 2015 |
| Last update date |
Aug 31, 2016 |
| Contact name |
Akira Higashibata |
| E-mail(s) |
[email protected]
|
| Phone |
050-3362-3898
|
| Organization name |
Japan Aerospace Exploration Agency
|
| Department |
Department of Space Biology and Microgravity Sciences
|
| Lab |
ISS Science Project office
|
| Street address |
2-1-1,Sengen
|
| City |
Tsukuba |
| State/province |
Ibaraki |
| ZIP/Postal code |
305-8505 |
| Country |
Japan |
| |
|
| Platform ID |
GPL11346 |
| Series (1) |
| GSE71770 |
Microgravity effect on C. elegans N2/VC (CERISE, 4 days) |
|
