期刊
DEVELOPMENTAL DYNAMICS
卷 238, 期 12, 页码 3168-3174出版社
WILEY
DOI: 10.1002/dvdy.22143
关键词
melting curve; zebrafish; genotyping; SNP; point mutant; insertional mutant
资金
- National Heart, Lung
- Blood Institute [HL66292]
- NIH [GM082116]
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(mer) and p53(zy7) (P53(I166T)), a small deletion mutant (hap28(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. Developmental Dynamics 238:3168-3174, 2009. (C) 2009 Wiley-Liss, Inc.
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