|
| Status |
Public on Jul 23, 2010 |
| Title |
Ex3aERKO Uterus_estradiol_24h_rep2 |
| Sample type |
RNA |
| |
|
| Source name |
Ex3aERKO uterus, E2 treated 24 hours
|
| Organism |
Mus musculus |
| Characteristics |
esr1 genotype: Ex3aERKO (Esr1 null)
agent: E2
time point: 24h
strain: C57bl6/Jx129
age: adult (10+ weeks)
gender: ovariectomized female
tissue: uterus
|
| Treatment protocol |
ovariectomized mice were allowed to rest for 10-14 days, then injected with 100 ul saline or 250 ng estradiol in 100 ul saline ip. Uterine tissue was collected 2 or 24 hours after injection and frozen in liquid nitrogen
|
| Extracted molecule |
total RNA |
| Extraction protocol |
frozen uterine tissue was pulverized and each uterus was homogenized in 800 ul of Trizol (Invitrogen) with 250 ug/ml glycogen. Extraction proceeded as descibed in manufacturer's protocol. Following resuspension of RNA pellet, a second preciptation was done with NaAcetate and ethanol. This pellet was then resuspended in depc treated water and cleaned up using the Qiagen RNeasy kit clean up protocol. RNA concentration and A260/280 was checked and quality was evaluated by formaldehyde gel.
|
| Label |
Cy3
|
| Label protocol |
Starting with 500ng of total RNA, Cy3 labeled cRNA was produced according to manufacturer’s protocol.
|
| |
|
| Hybridization protocol |
For each sample, 1.65ug of Cy3 labeled cRNAs were fragmented and hybridized for 17 hours in a rotating hybridization oven.
|
| Scan protocol |
Slides were washed and then scanned with an Agilent Scanner
|
| Description |
gene expression in Ex3aERKO (ERa null) ovariectomized E2 24 hour treated uterus
|
| Data processing |
Data was obtained using the Agilent Feature Extraction software (v9.5), using the 1-color defaults for all parameters. The Agilent Feature Extraction Software performed error modeling, adjusting for additive and multiplicative noise. The default setting for the protocol we used is polynomial data fit.
|
| |
|
| Submission date |
Jul 22, 2010 |
| Last update date |
Jul 22, 2010 |
| Contact name |
NIEHS Microarray Core |
| E-mail(s) |
microarray@niehs.nih.gov, liuliw@niehs.nih.gov
|
| Organization name |
NIEHS
|
| Department |
DIR
|
| Lab |
Microarray Core
|
| Street address |
111 T.W. Alexander Drive
|
| City |
RTP |
| State/province |
NC |
| ZIP/Postal code |
27709 |
| Country |
USA |
| |
|
| Platform ID |
GPL4134 |
| Series (1) |
| GSE23072 |
Estrogen response uterine gene profile in Ex3αERKO |
|
