|
| Status |
Public on Dec 07, 2012 |
| Title |
d11.5 placenta leptin 29 |
| Sample type |
RNA |
| |
|
| Source name |
d11.5 placenta, mom 151
|
| Organism |
Mus musculus |
| Characteristics |
treatment: 50% food restriction w/ leptin replacement
tissue: placenta
maternal treatment group: leptin
pregnancy stage: d11.5
genetic background: Swiss Webster
|
| Treatment protocol |
Mice were mated, and on pregnancy day 1.5, randomized to (1) ad libitum fed (control); (2) restricted to 50% of previous food consumption (restricted); (3) restricted to 50% and injected once daily with 1 mg/kg recombinant mouse leptin. Mice were sacrificed on d11.5 and placentas were collected.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Placentas were halved and frozen in Trireagent (Sigma). After phase separation, the aqueous phase was applied to RNEasy columns (Qiagen) according to manufacturer's protocol. Total RNA from two placentas per mother were pooled.
|
| Label |
Cy3
|
| Label protocol |
First and second strand cDNA was prepared from the total RNA samples. cRNA target was prepared from the DNA template and verified on the Bioanalyzer.
|
| |
|
| Hybridization protocol |
Purified cRNA (1 µg) was fragmented to uniform size and applied to Agilent 4x44K Whole Mouse Genome microarrays (Design ID 026655, Agilent Technologies, Santa Clara, CA) in hybridization buffer. Arrays were hybridized at 65° C for 17 h on a rotating incubator and washed at 37° C for 1 min.
|
| Scan protocol |
Rinsed and dried arrays were scanned with an Agilent G2565 Microarray Scanner (Agilent Technologies, Santa Clara, CA) at 5 µm resolution.
|
| Description |
pooled from two placentas
|
| Data processing |
Agilent Feature Extraction software was used to process the scanned images from arrays (gridding and feature intensity extraction) and the data generated for each probe on the array was analyzed with GeneSpring GX v7.3.1 software (Agilent Technologies, Santa Clara, CA).
|
| |
|
| Submission date |
May 21, 2012 |
| Last update date |
Dec 07, 2012 |
| Contact name |
Laura Clamon Schulz |
| E-mail(s) |
[email protected]
|
| Organization name |
University of Missouri
|
| Department |
Obstetrics, Gynecology and Women's Health
|
| Street address |
1 Hospital Dr.
|
| City |
Columbia |
| State/province |
MO |
| ZIP/Postal code |
65212 |
| Country |
USA |
| |
|
| Platform ID |
GPL11202 |
| Series (1) |
| GSE38095 |
Leptin and the Placental Response to Maternal Food Restriction during Early Pregnancy |
|
