At different developmental time points we isolate discrete elements of the kidney by using laser capture microdissection and then define their gene expression profiles with microarrays.
Array Hybridization
Molecule:
Total RNA
A_260_280:
NA
RNA Extraction Protocol:
Potter protocol: 'RNA purification'
Target Amplification Manufacturer/kit:
Epicentre
Target Amplification Protocol:
Potter protocols
Rounds of Amplification:
2
Amount Labeled Target Hybridized To Array:
5ug
Label:
Biotin
Array Hyb/Wash Protocol:
Affymetrix standard protocol
GCOS Tgt Value:
1500
Data Analysis Method:
Affymetrix GCOS and Gene Spring and Avadis programs
Gene expression profiles of components of the developing kidney
Summary:
The long term objective is to create an encyclopedia of the expression levels of all genes in multiple components of the developing kidney. The central thesis is straightforward. The combination of 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 kidney. Microarrays with essentially complete genome coverage can be used to quantitate expression levels of every gene in laser capture microdissected components of the developing kidney. 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.
Type:
Comparison of kidney components.
Overall Design:
At different developmental time points we isolate discrete elements of the kidney by using laser capture microdissection and then define their gene expression profiles with microarrays.
