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A rapid and efficient method of genotyping zebrafish mutants

✍ Scribed by John M. Parant; Stephen A. George; Rob Pryor; Carl T. Wittwer; H. Joseph Yost

Publisher
John Wiley and Sons
Year
2009
Tongue
English
Weight
441 KB
Volume
238
Category
Article
ISSN
1058-8388

No coin nor oath required. For personal study only.

✦ Synopsis

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. Developmental Dynamics 238:3168–3174, 2009. © 2009 Wiley‐Liss, Inc.

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