|
| Status |
Public on Sep 14, 2017 |
| Title |
mammary epithelial cells from lactating sow, 3 h post challenge with heat-inactivated E. coli [918_Pool_3 hpc] |
| Sample type |
RNA |
| |
|
| Source name |
miRNA expression data from mammary epithelial cells of lactating sow 3 hpc with E. coli
|
| Organism |
Sus scrofa |
| Characteristics |
developemental stage: lactating sow
cell type: porcine mammary epithelial cells
challenged with: heat-inactivated E. coli
time point: 3 h post challenge
|
| Treatment protocol |
Mammary epithelial cells from lactating sows were challenged for 3 h and for 24 h with heat-inactivated E. coli isolate from milk of PDS-positive sows. Unchallenged cells were carried as controls.
|
| Growth protocol |
Approximately 4.4 x 10^5 cells from each individual were seeded and cultured in collagen-coated 6-well plates. Cells were grown to a confluency of about 90% and then experiment started.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated using Qiagen miReasy Mini kit TRI Reagent (Qiagen, Hilden, Germany) according to the manufacturer’s protocol for small-RNA. Then small RNA was cleaned up using the Rneasy MinElute Clean up kit (Qiagen, Hilden, Germany).
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared from miRNA according to FlashTag Biotin RNA Labeling Kit for Affymetrix GeneChip miRNA arrays (Genisphere, Hatfield, PA, USA) according to the manufacturer’s recommendations Affymetrix GeneChip miRNA Array protocols (Affymetrix, Santa Clara, CA, USA)
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 ug of cRNA were hybridized for 16 hr at 42°C on Affymetrix GeneChip miRNA arrays. GeneChips were washed and processed to detect biotin-containing transcripts by Streptavidin-PE (Phycoerythrin) conjugate.
|
| Scan protocol |
Scanning using the GeneChip scanner 3000 were performed according to Affymetrix protocols
|
| Description |
918_Pool_3 hpc
miRNA expression data from mammary epithelial cells of lactating sow 3 hpc with E. coli
|
| Data processing |
Data were analysed with Expression Console and quantified by med polish
|
| |
|
| Submission date |
Oct 18, 2016 |
| Last update date |
Sep 14, 2017 |
| Contact name |
Siriluck Ponsuksili |
| E-mail(s) |
[email protected]
|
| Organization name |
FBN
|
| Street address |
Wilhelm-stahl-allee 2
|
| City |
Dummerstorf |
| ZIP/Postal code |
18196 |
| Country |
Germany |
| |
|
| Platform ID |
GPL16384 |
| Series (1) |
| GSE88870 |
MicroRNA expression profiling of porcine mammary epithelial cells after challenge with Escherichia coli in vitro |
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