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Yuko Nishiwaki
  • Developmental neurobiology unit, Okinawa Institute of Science and Technology Graduate University, Japan
PCR-Based Genotyping of Zebrafish Genetic Mutants
基于 PCR 的斑马鱼基因突变体分型方法
作者:Swathy Babu, Yuko Nishiwaki and Ichiro Masai日期:03/20/2025,浏览量:485,Q&A: 0

Zebrafish genetic mutants have emerged as a valuable model system for studying various aspects of disease and developmental biology. Mutant zebrafish embryos are generally identified based on phenotypic defects at later developmental stages, making it difficult to investigate underlying molecular mechanisms at earlier stages. This protocol presents a PCR-based genotyping method that enables the identification of wild-type, heterozygous, and homozygous zebrafish genetic mutants at any developmental stage, even when they are phenotypically indistinguishable. The approach involves the amplification of specific genomic regions using carefully designed primers, followed by gel electrophoresis. This genotyping method facilitates the investigation of the molecular mechanisms driving phenotypic defects that are observed at later timepoints. This protocol allows researchers to perform analyses such as immunofluorescence, RT-PCR, RNA sequencing, and other molecular experiments on early developmental stages of mutants. The availability of this protocol expands the utility of zebrafish genetic mutants for elucidating the molecular underpinnings of various biological processes throughout development.