|
| Status |
Public on Mar 01, 2010 |
| Title |
WT Placenta_Saline_rep3 |
| Sample type |
RNA |
| |
|
| Source name |
Placenta harvested from WT fetuses injected in utero with saline
|
| Organism |
Mus musculus |
| Characteristics |
tissue: Placenta
genotype: WT
embryonic day: 18.5
gender: female
treatment: saline
|
| Treatment protocol |
Pregnant dams were anesthetized, a pelvic incision was made, and fetuses were given an intraabdominal injection of 1 nmol of PTH (1-84) in 2µl of saline or 2 µl of saline. 90 minutes later placenta were collected, snap-frozen in liquid nitrogen, and stored at – 70 °C for subsequent RNA extraction.
|
| Growth protocol |
Adult mice were given water and a standard chow (1% calcium) ad lib. Female heterozygous-deleted mice were subjected to timed matings with heterozygous-deleted stud males, allowing placenta to be consistently harvested on ED 18.5.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using the RNeasy Midi Kit (Quiagen). Extraction was performed as per the product insert.
|
| Label |
Biotin
|
| Label protocol |
Biotinylated cRNA were prepared from 200 ng of total RNA. This was preformed using the Affymetrix GeneChip Whole Transcript (WT) Sense Target Labeling Assay, and was carried out as per the user manual.
|
| |
|
| Hybridization protocol |
Following fragmentation and labeling, 5.5 ug of cRNA were hybridized on the GeneChip Mouse Gene 1.0 ST Array. GeneChips were then washed and stained according to the FS450_007 fluidics protocol. The Affymetrix GeneChip Hybridization, Wash, and Staining Kit was utilized, and protocols were carried out as per the users manual.
|
| Scan protocol |
GeneChips were scanned in the Affymetrix GeneChip Scanner 3000.
|
| Description |
Fetuses were treated in utero with saline, 90 min prior to harvesting the placenta.
|
| Data processing |
The data were normalized with RMA algorithm implemented in bioconductor (www.bioconductor.org). Probesets without gene names/gene assignments were removed from further analyses.
|
| |
|
| Submission date |
Jul 07, 2009 |
| Last update date |
Feb 26, 2010 |
| Contact name |
Christopher S Kovacs |
| E-mail(s) |
ckovacs@mun.ca
|
| Phone |
1-709-777-6881
|
| Fax |
1-709-777-8049
|
| URL |
http://www.med.mun.ca/Medicine/Faculty/Kovacs.aspx
|
| Organization name |
Memorial University of Newfoundland
|
| Department |
Faculty of Medicine - Endocrinology
|
| Lab |
Calcium Research Laboratory
|
| Street address |
Health Sciences Centre, 300 Prince Philip Drive
|
| City |
St. John's |
| State/province |
Newfoundland |
| ZIP/Postal code |
A1B 3V6 |
| Country |
Canada |
| |
|
| Platform ID |
GPL6246 |
| Series (1) |
| GSE16983 |
Expression data from placenta harvested from WT and Pth-null fetuses treated 90 minutes prior with saline or PTH (1-84) |
|
