|
| Status |
Public on Oct 30, 2014 |
| Title |
Human term placenta, intervention - male, biological replicate 5 |
| Sample type |
RNA |
| |
|
| Source name |
Human term placenta from women of the n-3 LCPUFA intervention group with male offspring
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: term placenta - villous fraction
group: n-3 LCPUFA intervention
offspring sex: male
|
| Treatment protocol |
After birth placentas were maintained and processed at 4-0°C. Placental samples were dissected from each of the four quadrants with the same distance from to the placenta center according to a standardized sampling protocol. To obtain the chorionic villous fraction of the placenta, maternal basal plate and fetal chorionic plate of each placenta sample were removed, avoiding large vessels, calcifications or other obvious abnormalities. The samples were snap-frozen and stored at -80°C until RNA extraction.
|
| Growth protocol |
-
|
| Extracted molecule |
total RNA |
| Extraction protocol |
For each placenta, placental villous fraction samples from the four quadrants were homogenised in TRIzol® Reagent (Invitrogen, Darmstadt, Germany) and pooled in equal parts for total RNA extraction by midi RNeasy Kit (Qiagen, Hilden, Germany), each according to the manusfacturer's instructions
|
| Label |
biotin
|
| Label protocol |
Per placenta, 5ug total RNA was reverse transcribed to cDNA by Gene Chip Expression 3'-Aplification one cycle cDNA Synthesis Kit. cDNA was in-vitro transcribed to biotinylated cRNAs by Chip Expression 3'-Amplification IVT labelling Kit according to the Affymetrix's GeneChip Expression Analysis Technical manual.
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 ug of cRNA were hybridized for 16 hr at 45°C on Affymetrix Custom Array - NuGO_Hs1a520180 array. GeneChips were washed and stained in the Affymetrix Fluidics Station 400.
|
| Scan protocol |
GeneChips were scanned using the GeneChip Scanner 3000.
|
| Description |
Placenta INT male 5
|
| Data processing |
MADMAX (Management and analysis Database for Multi-platform microArray eXperiments) was applied for quality control, normalization (gc Robust Multichip Average (slow)) and statistical analysis. Custom CDF file 13.0.0 were used.
|
| |
|
| Submission date |
Dec 13, 2013 |
| Last update date |
Oct 31, 2014 |
| Contact name |
Bernhard L Bader |
| Organization name |
Technische Universität München
|
| Department |
Clinical Nutritional Unit - ZIEL-Research Center for Nutrition and Food Sciences
|
| Lab |
ZIEL-PhD-Group ‘Epigenetics, Imprinting and Nutrition’
|
| Street address |
Gregor-Mendel Straße 2
|
| City |
Freising-Weihenstephan |
| ZIP/Postal code |
85350 |
| Country |
Germany |
| |
|
| Platform ID |
GPL18060 |
| Series (1) |
| GSE53291 |
Sex-specific changes of placental transcriptome in response to a n-3 LCPUFA supplementation of pregnant women |
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