|
| Status |
Public on May 29, 2019 |
| Title |
ECO_RAVI_2W_1 |
| Sample type |
RNA |
| |
|
| Source name |
ECO_RAVI_2W_1
|
| Organism |
Sus scrofa |
| Characteristics |
tissue: whole blood
|
| Treatment protocol |
Two weeks after the stocking, the porcine atropic thinitis vaccine was administered pigs at 2 weeks, 4 weeks, and 8 weeks post vaccination.
|
| Growth protocol |
Twenty of six-week-old waened pigs were raised in two rooms, each equipped with a gasoline heater and a mineral heat ware for 8 weeks.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Tota RNA was isolated form the experimental porcine using PAXgene Blood RNA Kit (Qiagen, Valencia, CA, USA) according to the manufacturer's protocol.
|
| Label |
Cy3
|
| Label protocol |
RNA labeling and hybridization were performed by using the Agilent One-Color Microarray-Based Gene Expression Analysis protocol (Agilent Technology, V 6.5 2010). Briefly, 100ng of total RNA form each sample was linearly amplified and labeled with Cy3-dCTP. The labeled cRNAs were purified by RNeasy Mini Kit (Qiagen). The concentration and specific activity of the labeled cRNAs (pmol Cy3/ug cRNA) were measured by NanoDrop ND-1000 (NanoDrop, Wilmington, USA).
|
| |
|
| Hybridization protocol |
1650ng of each labeled cRNA was fragmented by adding 11 µl 10x blocking agent and 2.2 µl of 25 x fragmentation buffer, and then heated at 60℃ for 30 min. Finally, 55 ul 2 x GE hybridization buffer was added to dilute the labeled cRNA. 100 µl of hybridization solution was dispensed into the gasket slide and assembled to the Agilent SurePrint HD Porcine GE 4X44K Microarrays (Agilent®). The slides were incubated for 17 h at 65 ℃ in an Agilent hybridization oven. Then washed at room temperature by using the Agilent One-Color Microarray-Based Gene Expression Analysis protocol (Agilent Technology, V 6.5, 2010).
|
| Scan protocol |
The hybridized array was immediately scanned with an Agilent Microarray Scanner D (Agilent Technologies, Inc.)
|
| Description |
gene expression of porcine whole blood raised in the RAVI system for 2 weeks
|
| Data processing |
Microarray results were extracted using Agilent Feature Extraction software v11.0 (Agilent Technologies). Array probes that have Flag A in samples were filtered out.
|
| |
|
| Submission date |
May 28, 2019 |
| Last update date |
May 29, 2019 |
| Contact name |
Suji Kim |
| E-mail(s) |
[email protected]
|
| Organization name |
Seoul National University
|
| Department |
Veterinary Medicine
|
| Lab |
Infectious Disease
|
| Street address |
1, Gwanak-ro
|
| City |
Seoul |
| State/province |
Gwanak-gu |
| ZIP/Postal code |
ASI/KR/KS013 |
| Country |
South Korea |
| |
|
| Platform ID |
GPL16571 |
| Series (1) |
| GSE131821 |
Evaluation of immunostimulating effects of mineral heat ware in pig |
|
