A rapid and efficient method of genotyping zebrafish mutants
- Parant, J.M., George, S.A., Pryor, R., Wittwer, C.T., and Yost, H.J.
- Developmental Dynamics : an official publication of the American Association of Anatomists 238(12): 3168-3174 (Journal)
- Registered Authors
- Parant, John, Yost, H. Joseph
- melting curve, zebrafish, genotyping, SNP, point mutant, insertional mutant
- MeSH Terms
- Animals, Genetically Modified
- Base Sequence
- Cytogenetic Analysis/methods*
- DNA Mutational Analysis/methods*
- Embryo, Nonmammalian
- Gene Deletion
- Molecular Sequence Data
- Mutagenesis, Insertional
- Mutant Proteins/analysis
- Mutant Proteins/genetics
- Polymorphism, Single Nucleotide
- Time Factors
- Zebrafish/growth & development
- 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.
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.
Genes / Markers
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Engineered Foreign Genes