|ZFIN ID: ZDB-PUB-091120-24|
A rapid and efficient method of genotyping zebrafish mutants
Parant, J.M., George, S.A., Pryor, R., Wittwer, C.T., and Yost, H.J.
|Source:||Developmental dynamics : an official publication of the American Association of Anatomists 238(12): 3168-3174 (Journal)|
|Registered Authors:||Parant, John, Yost, H. Joseph|
|Keywords:||melting curve, zebrafish, genotyping, SNP, point mutant, insertional mutant|
|PubMed:||19890916 Full text @ Dev. Dyn.|
Parant, J.M., George, S.A., Pryor, R., Wittwer, C.T., and Yost, H.J. (2009) A rapid and efficient method of genotyping zebrafish mutants. Developmental dynamics : an official publication of the American Association of Anatomists. 238(12):3168-3174.
ABSTRACTIn 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.