Quantitative genotyping of mouse brain-specific PEX13 gene disruption by real-time PCR

Müller, C. Catharina, Nourse, Jamie P., Nguyen, Tam H. and Crane, Denis I. (2009) Quantitative genotyping of mouse brain-specific PEX13 gene disruption by real-time PCR. Journal of Neuroscience Methods, 181 1: 73-81. doi:10.1016/j.jneumeth.2009.04.025


Author Müller, C. Catharina
Nourse, Jamie P.
Nguyen, Tam H.
Crane, Denis I.
Title Quantitative genotyping of mouse brain-specific PEX13 gene disruption by real-time PCR
Journal name Journal of Neuroscience Methods   Check publisher's open access policy
ISSN 0165-0270
Publication date 2009-06-30
Sub-type Article (original research)
DOI 10.1016/j.jneumeth.2009.04.025
Volume 181
Issue 1
Start page 73
End page 81
Total pages 9
Place of publication The Netherlands
Publisher Elsevier BV
Language eng
Subject 1109 Neurosciences
1702 Cognitive Sciences
Formatted abstract
The Cre/loxP-system has become an invaluable tool for the generation of tissue-specific gene disruption in mice. However, because Cre recombinase excision of individual genes can be variable, an accurate and sensitive method is necessary to determine the ultimate level of gene disruption. The analysis of gene disruption is particularly difficult for tissue that has been fixed for (immuno)histochemical analysis with paraformaldehyde. Here, we describe a simple, rapid and cost effective method for measurement of gene disruption using quantitative real-time PCR, through application to the analysis of PEX13 gene disruption in a brain-specific PEX13 mouse mutant. We show that this general protocol is suitable for both normal and paraformaldehyde-fixed tissue.
Keyword Quantitative real-time PCR
Genotyping
Gene excision efficiency
Cre recombinase
Tissue-specific gene disruption
Conditional mutants
Fixed tissue
Transgenic mice
Q-Index Code C1
Q-Index Status Provisional Code

Document type: Journal Article
Sub-type: Article (original research)
Collection: Queensland Brain Institute Publications
 
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Created: Fri, 24 Sep 2010, 15:50:17 EST by Laura McTaggart on behalf of Queensland Brain Institute