|
| Status |
Public on Dec 01, 2009 |
| Title |
E2_2h vs. WT V |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
WT uterus, E2, 2h
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL/6
gender: female
age: 10+ weeks
tissue: uterus
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol with Qiagen RNA easy clean up
|
| Label |
Cy3
|
| Label protocol |
Total RNA was amplified using the Agilent Low RNA Input Fluorescent Linear Amplification Kit protocol. Starting with 500ng of total RNA, Cy5 labeled cRNA was produced according to manufacturer’s protocol.
|
| |
|
| Channel 2 |
| Source name |
WT uterus, vehicle, 2h
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL/6
gender: female
age: 10+ weeks
tissue: uterus
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol with Qiagen RNA easy clean up
|
| Label |
Cy5
|
| Label protocol |
Total RNA was amplified using the Agilent Low RNA Input Fluorescent Linear Amplification Kit protocol. Starting with 500ng of total RNA, Cy3 labeled cRNA was produced according to manufacturer’s protocol.
|
| |
|
| |
| Hybridization protocol |
For each two color comparison, 750ng of each Cy3 and Cy5 labeled cRNAs were mixed and fragmented using the Agilent In Situ Hybridization Kit protocol. Hybridizations were performed for 17 hours in a rotating hybridization oven using the Agilent 60-mer oligo microarray processing protocol.
|
| Scan protocol |
Slides were washed as indicated in the Agilent 60-mer oligo microarray processing protocol and then scanned with an Agilent Scanner. Data was obtained using the Agilent Feature Extraction software, using defaults for all parameters.
|
| Description |
pooled RNA from three vehicle or three treated uteri
|
| Data processing |
Data was obtained using the Agilent Feature Extraction software (v8.1), using defaults for all parameters and loaded into the Rosetta Resolver® system (v6.0) (Rosetta Biosoftware, Kirkland, WA).
|
| |
|
| Submission date |
Sep 17, 2009 |
| Last update date |
Sep 22, 2009 |
| Contact name |
NIEHS Microarray Core |
| E-mail(s) |
[email protected], [email protected]
|
| 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 |
GPL7202 |
| Series (1) |
| GSE18168 |
Uterine gene profiles from WT, KIKO (DNA-binding deficient ERα) and aERKO mice treated with estradiol or xenoestrogens |
|
