|
| Status |
Public on Mar 23, 2023 |
| Title |
MCTWC1 |
| Sample type |
RNA |
| |
|
| Source name |
frontal cortexes
|
| Organism |
Mus musculus |
| Characteristics |
stain: CD-1
gender: Male
|
| Treatment protocol |
Detection of a vaginal plug was considered day 0.5 of pregnancy. Pregnant CD-1 mice were then housed individually and continuously treated with E3 (50µl of 0.46mg/ml in DMSO to approximate human exposure) or vehicle control (DMSO) via a mini-osmotic pump (Alzet model # 1002) beginning on Day 9 of gestation for 7 days.We began each experiment by treating pregnant mice with E3, and ended by analyzing their offspring.
|
| Growth protocol |
CD-1 mice were obtained from Charles River Laboratories (Wilmington, MA, USA). Animals were housed in a 12 hr day/night cycle. All experiments were conducted in accordance with Yale University Animal Care Committee Guidelines. Eight-week-old CD-1 female mice were bred to male mice of the same strain and examined every 12 hours until the presence of a vaginal plug was detected.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
The tissue intended for mRNA analysis was placed in 1 mL TRIzol per 100 mg tissue (Invitrogen, Carlsbad, CA, USA). Tissue was homogenized on ice, and total RNA was isolated using methodology described by the manufacturer.
|
| Label |
biotin
|
| Label protocol |
High quality total RNA is used as starting material to obtain labeled cRNA. In the first step, single stranded cDNA is synthesized by reverse transcription using the poly (A) RNA present in the starting total RNA sample. Single stranded cDNA is then converted into double stranded cDNA and purified using the Affymetrix Cleanup Module. An in vitro transcription (IVT) reaction is then carried out overnight in the presence of biotinylated
|
| |
|
| Hybridization protocol |
Samples are hybridized to arrays for 16 hours at 45C. The standard array is then washed and stained using the fluidics station and then scanned. The images are analyzed using Affymetrix Command Console Viewer and metric analyses are carried out according to the instructions provided by Affymetrix using the Affymetrix Expression Console.
|
| Scan protocol |
GeneChips were scanned using GeneChip Scanner 3000 with Autoloader.
|
| Data processing |
The data were analyzed with Expression Console using the following paramaters:
affymetrix-algorithm-name = sst-rma-gene-full
affymetrix-algorithm-version = 1.0
affymetrix-array-type = MTA-1_0
program-version = 1.4.1.46
|
| |
|
| Submission date |
Mar 22, 2022 |
| Last update date |
Mar 24, 2023 |
| Contact name |
Yuping Zhou |
| E-mail(s) |
joezhouyp@gmail.com
|
| Organization name |
Yale
|
| Department |
OB/GYN
|
| Lab |
Taylor Lab
|
| Street address |
310 cedar street
|
| City |
new haven |
| State/province |
CT |
| ZIP/Postal code |
06511 |
| Country |
USA |
| |
|
| Platform ID |
GPL20775 |
| Series (2) |
| GSE199174 |
The steroid hormone estriol (E3) regulates epigenetic programming of fetal mouse brain and reproductive tract [Affymetrix] |
| GSE199214 |
The steroid hormone estriol (E3) regulates epigenetic programming of fetal mouse brain and reproductive tract |
|
