|
| Status |
Public on Jan 10, 2018 |
| Title |
CD3+CD8+_rep3 |
| Sample type |
RNA |
| |
|
| Source name |
sow_PBMC_CD3+CD8+
|
| Organism |
Sus scrofa |
| Characteristics |
tissue: Whole blood
cell type: peripheral blood mononuclear cells (PBMC)
cell subtype: sorted lymphocyte
sorted fraction: CD3+CD8+
|
| Growth protocol |
PBMC was isolated by density gradient centrifugation on Percoll (d = 1.084) . CD3+CD4+, CD3+CD8+ and CD3+CD4+CD8+ were separately sorted by a cell sorter.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions and purified by RNeasy MinElute Cleanup kit.
|
| Label |
Cy3
|
| Label protocol |
Complementary DNA amplification was carried out from 25 ng of total RNA using an Ovation PicoSL WTA System V2. After amplification, cDNA samples were labeled with Cy3 using Klenow polymerase.
|
| |
|
| Hybridization protocol |
Slide was hybridized with 1.65 μg Cy3-labeled cDNA using a Gene Expression Hybridization Kit (Agilent technologies, Santa Clara, CA, USA) in a hybridization oven.
|
| Scan protocol |
After 16 h of hybridization at 65 °C, the slide was washed with a Gene Expression Wash Buffer Kit (Agilent technologies, Santa Clara, CA, USA), according to the manufacturer’s instructions. Slides were scanned after washing on the Agilent DNA Microarray Scanner (G2505C) using one color scan setting for 4x44k array slides.
|
| Description |
CD8#3
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 11.0.1.1 (Agilent) using default parameters to obtain background subtracted and spatially detrended Processed Signal intensities. The intensity signals was imported to CLC Main workbench 6.0 and normalized by the quartile normalization method and log2 transformed.
|
| |
|
| Submission date |
Oct 23, 2017 |
| Last update date |
Jan 23, 2018 |
| Contact name |
Ryo Inoue |
| E-mail(s) |
[email protected]
|
| Organization name |
Kyoto Prefectural University
|
| Street address |
1-5, Hangi-cho, shimogamo
|
| City |
Kyoto |
| ZIP/Postal code |
606-8522 |
| Country |
Japan |
| |
|
| Platform ID |
GPL16571 |
| Series (1) |
| GSE105784 |
Gene expression in peripheral blood Tcells from sows |
|
