|
| Status |
Public on Dec 01, 2013 |
| Title |
Optimal Dose 5 |
| Sample type |
RNA |
| |
|
| Source name |
Tetramer-sorted M1-specific T cells, optimal dose
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: CD8+ T cells
treatment: influenza M1 peptide-pulsed monocyte-derived dendritic cells
peptide dosage: optimal
|
| Treatment protocol |
Different peptide doses were used to pulse dendritic cells (optimal dose: 10nM; high dose: 10uM).
|
| Growth protocol |
CD8+ T cells were stimulated with M1 peptide-pulsed monocyte-derived dendritic cells in complete RPMI media supplemented with T cell growth factor.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
T cells were harvested on D14 and tetramer sorted before Trizol-based RNA extraction.
|
| Label |
Cy3
|
| Label protocol |
To produce Cy3-labeled cRNA, the RNA samples were amplified and labeled using the Agilent Low Input Quick Amp Labeling Kit (Agilent Technologies) following the manufacturer's protocol. Yields of cRNA and the dye-incorporation rate were measured with the ND-1000 Spectrophotometer.
|
| |
|
| Hybridization protocol |
The hybridization procedure was performed according to the Agilent 60-mer oligo microarray processing protocol using the Agilent Gene Expression Hybridization Kit (Agilent Technologies). Briefly, 600 ng Cy3-labeled fragmented cRNA in hybridization buffer was hybridized overnight (17 hours, 65 °C) to Agilent Whole Human Genome Oligo Microarrays 8x60K using Agilent's recommended hybridization chamber and oven.
|
| Scan protocol |
Fluorescence signals of the hybridized Agilent Microarrays were detected using Agilent's Microarray Scanner System (Agilent Technologies).
|
| Description |
OD5
Gene expression.
|
| Data processing |
The Agilent Feature Extraction Software (FES) was used to read out and process the microarray image files. The software determines feature intensities (including background subtraction), rejects outliers and calculates statistical confidences. The signal intensities were normalized by dividing the intensity values by their median.
|
| |
|
| Submission date |
Jul 30, 2012 |
| Last update date |
Dec 01, 2013 |
| Contact name |
Yen-Ling Chiu |
| Organization name |
Johns Hopkins University
|
| Street address |
733 N. Broadway, BRB 615
|
| City |
Baltimore |
| State/province |
MD |
| ZIP/Postal code |
21205 |
| Country |
USA |
| |
|
| Platform ID |
GPL13607 |
| Series (1) |
| GSE39761 |
CD8+ T cell gene expression after different antigen dose stimulations |
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