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J Vis Exp. 2014 Feb 5;(84):e51138. doi: 10.3791/51138.

Rapid and efficient zebrafish genotyping using PCR with high-resolution melt analysis.

Author information

1
Division of Pediatric Neurology, Department of Pediatrics, University of Utah School of Medicine; Department of Neurobiology and Anatomy, University of Utah School of Medicine; Interdepartmental Program in Neurosciences, University of Utah School of Medicine.
2
Division of Pediatric Neurology, Department of Pediatrics, University of Utah School of Medicine.
3
Mutation Generation and Detection Core, HSC Core Research Facility, University of Utah School of Medicine.
4
Division of Pediatric Neurology, Department of Pediatrics, University of Utah School of Medicine; Department of Neurobiology and Anatomy, University of Utah School of Medicine; Interdepartmental Program in Neurosciences, University of Utah School of Medicine; Department of Neurology, University of Utah School of Medicine; Joshua.Bonkowsky@hsc.utah.edu; jbonkowsky@gmail.com.

Abstract

Zebrafish is a powerful vertebrate model system for studying development, modeling disease, and performing drug screening. Recently a variety of genetic tools have been introduced, including multiple strategies for inducing mutations and generating transgenic lines. However, large-scale screening is limited by traditional genotyping methods, which are time-consuming and labor-intensive. Here we describe a technique to analyze zebrafish genotypes by PCR combined with high-resolution melting analysis (HRMA). This approach is rapid, sensitive, and inexpensive, with lower risk of contamination artifacts. Genotyping by PCR with HRMA can be used for embryos or adult fish, including in high-throughput screening protocols.

PMID:
24561516
PMCID:
PMC4116811
DOI:
10.3791/51138
[Indexed for MEDLINE]
Free PMC Article

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