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Status |
Public on Jun 21, 2012 |
Title |
Vero E6 SARS CoV DeltaE 15hpi rep2 |
Sample type |
RNA |
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Source name |
Vero E6 cells, infected with SARS-CoV-DeltaE, 15hpi
|
Organism |
Chlorocebus aethiops |
Characteristics |
infection: SARS CoV DeltaE cell line: Vero E6 induce ifnβ: NO hours post infection: 15 hpi
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Growth protocol |
Subconfluent monolayers (90% confluency) of Vero E6 and MA-104 cells were infected at multiplicity of infection (moi) of 0.01 or 2 with the viruses rSARS-CoV-ΔE, and rSARS-CoV or rSARS-CoV-MA15-ΔE and rSARS-CoV-MA15. Culture supernatants were collected at different hours post-infection (hpi) and virus titer was determined as previously described.
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Extracted molecule |
total RNA |
Extraction protocol |
Total RNA was prepared using the RNeasy Mini Kit (Qiagen) according to the manufacturer´s instructions. RNA was quantified and the RNA quality was assessed with a 2100 Bioanalyzer (Agilent Technologies).
|
Label |
biotin
|
Label protocol |
Biotinylated cRNA was prepared according to the standard Affymetrix protocol from 5 µg total RNA (Expression Analysis Technical Manual, 2001, Affymetrix).
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Hybridization protocol |
Following fragmentation, 10 µg of biotinylated cRNA were hybridized to Affymetrix GeneChips (Human Genome U133 plus 2.0) at 45ºC during 16 hours. Each sample was added to a hybridization solution containing 100mM 2-(N-morpholino)ethanesulfonic acid, 1 M Na+, 20mM of EDTA and 0,01% tween-20
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Scan protocol |
The arrays were stained with streptavidin-ficoeritrin and read at 1.56 μm in a GeneChip Scanner 3000 7G System (Affymetrix, Santa Clara, CA)
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Description |
Gene expression data from Vero E6 cells infected with SARS-CoV-DeltaE and collected at 15hpi
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Data processing |
GeneChip intensities were background-corrected, normalized and sumarized by RMA method, using the “Affy” package from Bioconductor.
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Submission date |
Jul 12, 2011 |
Last update date |
Jun 21, 2012 |
Contact name |
Juan Carlos Oliveros |
Organization name |
CNB, CSIC
|
Street address |
Darwin 3
|
City |
Cantoblanco |
State/province |
Madrid |
ZIP/Postal code |
28049 |
Country |
Spain |
|
|
Platform ID |
GPL570 |
Series (1) |
GSE30589 |
Severe acute respiratory syndrome coronavirus envelope protein regulates cell stress responses and apoptosis |
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