Expression Database

GUDMAP:6966

Theiler Stage

Images

Supplemental Data Files

CEL file:	JS_7_MOE430_2.CEL
CHP file:	JS_7_MOE430_2.CHP
RPT file:	JS_7_MOE430_2.RPT
TXT file:	JS_7_MOE430_2.TXT

Archive ID

Principal Investigator(s)

James Lessard, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, USA, james.lessard@cchmc.org

Submitted By

John C Szucsik, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, USA, john.szucsik@cchmc.org

Specimen Details

Sample GEO ID:

GSM143564

Sample Description:

embryonic day 14 urogenital sinus

Title:

JS_7

Component(s) Sampled:

urogenital sinus (EMAP:6689)

Organism:

Mus musculus

Strain:

FVB/N

Genotype:

Non-wild type Allele:
Gene		Actg2
MGI Symbol or lab name		Tg(Actg2-EGFP)Jll
Type		transgenic insertion
Notes:		PMID: 14713859

Sex:

Unknown

Development Age:

14 dpc

Theiler Stage:

Developmental Landmark:

Individual fingers visible in the anterior footplate with indentations between the toes which are not separated.

Pool Size:

N/A

Pooled Sample:

Dissection Method:

Urogenital sinus region microdissected based on an absence of SMGA/EGFP expression

Experimental Design:

Female FVB/N mice were time-mated to SMAA/EYFP transgenic males. Pregnant FVB/N mice were euthanized by standard carbon dioxide asphyxiation at 14 days post-coitus. Embryonic samples were euthanized by decapitation and tissues were micro-dissected, rapidly frozen and stored at -80ºC until RNA was extracted for gene expression analysis.

Array Hybridization

Extracted Molecule:	total RNA
A260:280 Ratio:	ND
RNA Extraction Protocol:	Qiagen: RNeasy
Target Amplification Manufacturer/kit:	TargetAmp 2-Round Aminoallyl-aRNA Amplification Kit 1.0 (EPICENTRE Biotechnologies)
Target Amplification Protocol:	Potter protocols
Rounds of Amplification:	2
Amount Labeled Target Hybridized To Array:	10 ug
Label:	Biotin
Label Protocol:	Biotin-X-X-NHS (EPICENTRE Biotechnologies)
Array Hyb/Wash Protocol:	Affymetrix standard protocol
Scan Protocol:
GCOS Tgt Value:	1500
Data Analysis Method:	Affymetrix GCOS and Gene Spring and Avadis programs
Reference:	day 1 mouse/Potter

Series Details

Series GEO ID:	GSE6223
Number of Samples:	13 samples
Title:	Gene expression profiles of E14 bladder, urogenital sinus and urethra isolated from SMGA (Actg2) transgenic mice. (GUDMAP Series ID: 6)
Summary:	The long term objective is to create an encyclopedia of the expression levels of all genes in multiple components of the developing bladder. The central thesis is straightforward. The combination ofmicrodissected and laser capture microdissection (LCM) plus microarray analysis offers a powerful, efficient and effective method for the creation of a global gene expression atlas of the developing urogenital system. Microarrays with essentially complete genome coverage can be used to quantitate expression levels of every gene. The ensuing rapid read-out provides an expression atlas that is more sensitive, more economical and more complete than would be possible by in situ hybridizations alone. The data submitted here represents the gene expression profiles of the embryonic day 14 bladder, urogenital sinus, and urethra.
Type:	Gene expression comparison from developing regions of the mouse embryonic day 14 urogenital system
Overall Design:	Bladder, urogenital sinus, and urethra regions from embryonic day 14 SMGA/EGFP animals were microdissected and total RNA isolated for gene expression analysis using the Affymetrix MOE430 microarray chip.

Platform Details

Platform GEO ID:	GPL1261
Title:	Affymetrix GeneChip Mouse Genome 430 2.0 Array
Distribution:	commercial
Technology:	in situ oligonucleotide
Organism:	Mus musculus
Manufacturer:	Affymetrix
Manufacturer Protocol:	N/A
Catalogue Number:	N/A