|
| Status |
Public on Sep 28, 2013 |
| Title |
slide2-array2 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Control
|
| Organism |
Sus scrofa |
| Characteristics |
developmental stage: Expanded blastocytes
dietary supplement: No
|
| Treatment protocol |
Three different type of experimental diets were given to gilts: 1) basal diet 0.2 mg/kg of natural Se and 2.3 mg/kg of vitamin B6 (pyridoxine ) and top-dressed with 50 g of ground corn without supplemental Se or pyridoxine; 2) basal diet plus0.3 mg/kg of feed of inorganic Se, as sodium selenite, and 10 mg/kg of feed of hydro-chloride pyridoxine; 3) similar condition as group 2 and replaced inorganic Se with 0.3 mg/kg of feed of organic Se as Se-enriched yeast.
|
| Growth protocol |
Sows were slaughtered 5 days after the bevahiour estrus and embryos were recovered.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
ArcturusW PicoPureW RNA Isolation Kit (Applied Biosystems, Carlsbad, CA, USA) was used for RNA extraction from EB with on column removal of DNA by DNase treatment using RNase-Free DNase kit according to the protocol from Qiagen
|
| Label |
Cy5
|
| Label protocol |
Antisense RNA (aRNA) was generated using RiboAmp HSPlus kit beforw labelling with Cy3 or Cy5 Fluorophore using ULS Fluorescent Labeling Kit from Kreatech. Labelling was done under ozone free environmental chamber.
|
| |
|
| Channel 2 |
| Source name |
Inorganic Se + B6
|
| Organism |
Sus scrofa |
| Characteristics |
developmental stage: Expanded blastocytes
dietary supplement: Inorganic Se + B6
|
| Treatment protocol |
Three different type of experimental diets were given to gilts: 1) basal diet 0.2 mg/kg of natural Se and 2.3 mg/kg of vitamin B6 (pyridoxine ) and top-dressed with 50 g of ground corn without supplemental Se or pyridoxine; 2) basal diet plus0.3 mg/kg of feed of inorganic Se, as sodium selenite, and 10 mg/kg of feed of hydro-chloride pyridoxine; 3) similar condition as group 2 and replaced inorganic Se with 0.3 mg/kg of feed of organic Se as Se-enriched yeast.
|
| Growth protocol |
Sows were slaughtered 5 days after the bevahiour estrus and embryos were recovered.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
ArcturusW PicoPureW RNA Isolation Kit (Applied Biosystems, Carlsbad, CA, USA) was used for RNA extraction from EB with on column removal of DNA by DNase treatment using RNase-Free DNase kit according to the protocol from Qiagen
|
| Label |
Cy3
|
| Label protocol |
Antisense RNA (aRNA) was generated using RiboAmp HSPlus kit beforw labelling with Cy3 or Cy5 Fluorophore using ULS Fluorescent Labeling Kit from Kreatech. Labelling was done under ozone free environmental chamber.
|
| |
|
| |
| Hybridization protocol |
Hybridization mixture was prepared together with Spike-In control according to Agilent Gene Expression Hybridization Kit 60-mer oligo microarray protocol version 4.0.
|
| Scan protocol |
Arrays were immediately scanned at 5 μm resolution after slides were washing and drying using an Axon 4200AL scanner (635 nm for Cy5 and at 532 nm for Cy3) using the autoscan feature from the default setting and images were analysed with Gene Pix Pro 6.0. software
|
| Description |
Dye-swap replicate of biological replicate 2
|
| Data processing |
FlexArray software was used for data normalization following first with simple background subtraction, LOWESS normalization within and between arrays.
|
| |
|
| Submission date |
Sep 27, 2013 |
| Last update date |
Sep 28, 2013 |
| Contact name |
Stephen Tsoi |
| E-mail(s) |
stsoi1@ualberta.ca
|
| Phone |
780-248-1567
|
| Organization name |
University of Alberta
|
| Department |
AFNS
|
| Street address |
4-32G Agriculture/Forestry Centre
|
| City |
Edmonton |
| ZIP/Postal code |
T6G 2P5 |
| Country |
Canada |
| |
|
| Platform ID |
GPL17779 |
| Series (1) |
| GSE51249 |
Impact of maternal diet supplemented with selenium (Se) and vitamin B6 during pregnancy on gene expression of porcine expanded blastocysts (EB) |
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