|
| Status |
Public on Mar 08, 2018 |
| Title |
Jurkat at 1g baseline, rep5 |
| Sample type |
RNA |
| |
|
| Source name |
Jurkat cells (ATCC Clone E6-1, TIB152™)
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: Jurkat T cells
|
| Treatment protocol |
Human Jurkat T cells were exposed for 5 min to simulated microgravity and hypergravity (9g) during ground based facility experiments.
|
| Growth protocol |
Jurkat T cells were cultivated in RPMI 1640 medium, supplemented with 10% fetal bovine serum, 2 mM glutamine and 100 U/ml penicillin as well as 100 µg/ml streptomycin. Cells were cultured with a density of 0.2x106 cells/ml and medium exchange was performed every 48 hours.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Jurkat cells were lysed with Trizol LS and total RNA was extracted following the Rneasy Midi Kit protocol (Qiagen).
|
| Label |
biotin
|
| Label protocol |
The fragmented and biotinylated DNA targets were prepared according to the standard Affymetrix WT PLUS Reagent Kit protocol (Affymetrix GeneChip® WT PLUS Reagent Kit, 902280) from 100 ng total RNA starting material and 5.5 µg cDNA intermediate product.
|
| |
|
| Hybridization protocol |
DNA targets were hybridized for 17h at 45°C on GeneChip Human Transcriptome Arrays 2.0. GeneChips were washed and stained in the Affymetrix Fluidics Station 450 according to the standard GeneChip Expression Wash, Stain and Scan protocol (Affymetrix GeneChip Wash, Stain and Scan Kit, 900720).
|
| Scan protocol |
Subsequently, the GeneChips were scanned using the Affymetrix 3000 7G scanner.
|
| Data processing |
For microarray data analysis, the Affymetrix Expression Console and Transcriptome Analysis Console was used. The robust multi-array (RMA) averaging method was applied for background correction, quantile normalization, and probe summarization.
|
| |
|
| Submission date |
Jul 10, 2017 |
| Last update date |
Mar 08, 2018 |
| Contact name |
Cora S. Thiel |
| E-mail(s) |
cora.thiel@uzh.ch
|
| Organization name |
University of Zurich
|
| Department |
Institute of Anatomy
|
| Street address |
Winterthurerstrasse 190
|
| City |
Zurich |
| ZIP/Postal code |
8057 |
| Country |
Switzerland |
| |
|
| Platform ID |
GPL17586 |
| Series (2) |
| GSE101101 |
Gene expression response to simulated gravity and hypergravity in human T cells |
| GSE101102 |
Gene expression response to altered gravity, simulated gravity and hypergravity in human T cells |
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