High-throughput DNA Extraction and Genotyping of 3dpf Zebrafish Larvae by Fin Clipping

Area of Science:

• Comparative Genomics
• Developmental Biology
• Genetics

Background:

• Zebrafish (Danio rerio) are valuable models for human disease research due to conserved gene orthologues.
• Genetic manipulation and disease modeling in zebrafish are advancing rapidly with gene editing tools.
• Current genotypic screening methods are often delayed until adulthood or postmortem, hindering early experimental design.

Purpose of the Study:

• To develop a rapid, cost-effective method for early genotype identification in zebrafish larvae.
• To enable precise selection of genetically modified zebrafish for downstream research.
• To facilitate high-throughput screening for experiments requiring known genotypes.

Main Methods:

• Adaptation of a protocol for isolating PCR-ready genomic DNA from live zebrafish larvae.
• Utilizing microscopic fin biopsies for DNA extraction at 72 hours post-fertilization (hpf).
• Implementation of a high-throughput PCR-based genotyping procedure.

Main Results:

• Successful isolation of PCR-ready genomic DNA from live zebrafish larvae as early as 72 hpf.
• Demonstration that fin biopsies are sufficient for accurate genotyping.
• Fins regenerate, allowing larvae to develop to adulthood after biopsy.

Conclusions:

• The developed technique provides a time- and cost-effective solution for early zebrafish genotyping.
• This method supports efficient selection and propagation of desired genotypes for research.
• Improved genotypic screening enhances the utility of zebrafish models in disease research and genetic studies.