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.
Date: 2009
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
MeSH Terms:
  • Animals
  • Animals, Genetically Modified
  • Base Sequence
  • Cytogenetic Analysis/methods*
  • DNA Mutational Analysis/methods*
  • Efficiency
  • Embryo, Nonmammalian
  • Gene Deletion
  • Genotype
  • Molecular Sequence Data
  • Mutagenesis, Insertional
  • Mutant Proteins/analysis
  • Mutant Proteins/genetics
  • Polymorphism, Single Nucleotide
  • Retroviridae/genetics
  • Retroviridae/physiology
  • Time Factors
  • Zebrafish/embryology
  • Zebrafish/genetics*
  • Zebrafish/growth & development
  • Zebrafish/metabolism
PubMed: 19890916 Full text @ Dev. Dyn.
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.
ADDITIONAL INFORMATION