TY  - JOUR
TI  - PCR-Based Genotyping of Zebrafish Genetic Mutants
AU  - Babu, Swathy 
AU  - Nishiwaki, Yuko 
AU  - Masai, Ichiro 
VL  - 15
IS  - 6
PY  - 2025
DA  - 2025/03/20
SP  - e5248
C1  - Bio-protocol 2025;15:e5248
DO  - 10.21769/BioProtoc.5248
UR  - https://doi.org/10.21769/BioProtoc.5248
AB  - 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.
KW  - Zebrafish
KW  - Genotyping
KW  - PCR
KW  - Mutant analysis
KW  - Developmental biology
KW  - Embryos
KW  - Restriction enzyme digestion
KW  - Phenotypic defects
KW  - Model organism
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False