|
| Status |
Public on Jan 26, 2015 |
| Title |
Endometrial inter-site_pregnant sows_day 18_rep3 |
| Sample type |
RNA |
| |
|
| Source name |
Endometrial inter-site, gestation day 18
|
| Organism |
Sus scrofa |
| Characteristics |
tissue: Endometrial inter-site
developmental stage: pregnant
time: day 18
breed: Large White
|
| Treatment protocol |
The sows of pregnant group were inseminated twice, 12 and 24 h after heat detection. The sows of non-pregnant group were treated with inactivate sperms from the same boar. Nine pregnant sows were slaughtered by electrical on day 13, day 18 and day 24 after insemination (the pregnant group, three sows every period). The non-pregnant sows were slaughtered on day 13 after inseminated (n=3).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol reagent (Invitrogen, Carlsbad, CA, USA) was used to extract total RNA according to the manufacturer’s instructions.
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 0.5 ug RNA using the One-Color Low RNA Input Linear Amplification PLUS kit (Agilent) according to the manufacturer's instructions, followed by RNAeasy column purification (QIAGEN, Valencia, CA). Dye incorporation and cRNA yield were checked with the NanoDrop ND-1000 Spectrophotometer.
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| |
|
| Hybridization protocol |
1.5 ug of Cy3-labelled cRNA (specific activity >10.0 pmol Cy3/ug cRNA) was fragmented at 60°C for 30 minutes in a reaction volume of 250 ml containing 1x Agilent fragmentation buffer and 2x Agilent blocking agent following the manufacturers instructions. On completion of the fragmentation reaction, 250 ml of 2x Agilent hybridization buffer was added to the fragmentation mixture and hybridized to Agilent Whole Human Genome Oligo Microarrays (G4112A) for 17 hours at 65°C in a rotating Agilent hybridization oven. After hybridization, microarrays were washed 1 minute at room temperature with GE Wash Buffer 1 (Agilent) and 1 minute with 37°C GE Wash buffer 2 (Agilent), then dried immediately by brief centrifugation.
|
| Scan protocol |
Slides were scanned immediately after washing on the Agilent DNA Microarray Scanner (G2505B) using one color scan setting for 1x44k array slides (Scan Area 61x21.6 mm, Scan resolution 10um, Dye channel is set to Green and Green PMT is set to 100%).
|
| Description |
Gene expression on day 18 of gestation
D18b-003
|
| Data processing |
The raw gene expression data were extracted from Agilent Feature Extraction Software (Version 10.7) and imported into Agilent GeneSpring GX software (version 11.0) for further analysis.
|
| |
|
| Submission date |
Oct 31, 2013 |
| Last update date |
Jan 26, 2015 |
| Contact name |
Xiao Chen |
| E-mail(s) |
[email protected]
|
| Organization name |
Institute of Apicultural Research, Chinese Academy of Agricultural Sciences
|
| Street address |
Haidian Xiangshan Beigou No.1
|
| City |
Beijing |
| State/province |
Beijing |
| ZIP/Postal code |
100093 |
| Country |
China |
| |
|
| Platform ID |
GPL16571 |
| Series (1) |
| GSE51970 |
Differential Gene Expression in Uterine Endometrium during Implantation in Pigs |
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