|
| Status |
Public on Nov 30, 2015 |
| Title |
Colostral CD8 single positive T cell replicate2 |
| Sample type |
RNA |
| |
|
| Source name |
Colostral CD8 single positive T cell from sow#2
|
| Organism |
Sus scrofa |
| Characteristics |
strain: Large white x Landrace
|
| Growth protocol |
colostrum and blood were collected from two Large White x Landrace sows. Mononuclear cells were isolated and fluorecently labeled by anti porcine CD3, CD4 and CD8 antibodies. CD3+/CD4-/CD8+ cells and CD3+/CD4+/CD8+ cells were separeted 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 |
Alexa Fluor 3
|
| Label protocol |
Complementary DNA amplification was carried out from 10 ng of total RNA using an Ovation PicoSL WTA System V2. After amplification, cDNA samples from colostrum were labeled with Alexa Fluor 3 and those from blood with Alexa Fluor 5 using a BioPrime Total Genomic Labeling System.
|
| |
|
| Hybridization protocol |
Slide was hybridized with 0.825 μg Cy3-labeled cDNA and 0.825 μg Cy5-labeled cDNA using a Gene Expression Hybridization Kit (Agilent technologies, Santa Clara, CA, USA) in a hybridization oven.
|
| Scan protocol |
After 17 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. The slide was then scanned by using GenePix 4100A (Axon, Foster City, CA).
|
| Description |
Biological replicate 2 of 2. Colostral CD8 single positive T cell
|
| Data processing |
The data was extracted with Axon GenePix Pro 7.2 image analysis software. The spot sizes and intensities quantified by the software and automatically flagged spot qualities were followed manual examination. Abnormal shape spots or spots with high local background or spots that were quantified due to false intensity caused by dust were flagged bad and discarded. The background subtracted median intensity signals was imported to CLC Main workbench 6.0 and normalized by the quartile normalization method and log2 transformed.
|
| |
|
| Submission date |
Aug 13, 2015 |
| Last update date |
Nov 30, 2015 |
| 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) |
| GSE72037 |
porcine T cells : colostral T cells vs blood T cells |
|
