A rapid and efficient method of genotyping zebrafish mutants.
Dev Dyn
; 238(12): 3168-74, 2009 Dec.
Article
em En
| MEDLINE
| ID: mdl-19890916
ABSTRACT
In order to facilitate high throughput genotyping of zebrafish, we have developed a novel technique that uses High Resolution Melting Analysis (HRMA) to distinguish wild-type, heterozygous mutants and homogyzous mutants. This one hour technique removes the need for restriction enzymes and agarose gels. The generated melting curve profiles are sensitive enough to detect non-specific PCR products. We have been able to reliably genotype three classes of mutations in zebrafish, including point mutants, apc(hu745) (apc(mcr)), and p53(zy7) (p53(I166T)), a small deletion mutant (bap28(y75)) and a retroviral insertion mutant (wdr43(hi821a)). This technique can genotype individual zebrafish embryos and adults (by tail-clip) and is applicable to other model organisms.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Peixe-Zebra
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Análise Mutacional de DNA
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Análise Citogenética
Idioma:
En
Ano de publicação:
2009
Tipo de documento:
Article