Rapid and Efficient Zebrafish Genotyping Using PCR with High-resolution Melt Analysis

Xing, Lingyan; Quist, Tyler S.; Stevenson, Tamara J.; Dahlem, Timothy J.; Bonkowsky, Joshua L.

doi:10.3791/51138

i

Rapid and Efficient Zebrafish Genotyping Using PCR with High-resolution Melt Analysis

Supporting Files

Feb 05 2014
By Xing, Lingyan ; Quist, Tyler S. ; Stevenson, Tamara J. ; ...

Details

Alternative Title:

J Vis Exp
Personal Author:

Xing, Lingyan ; Quist, Tyler S. ; Stevenson, Tamara J. ; Dahlem, Timothy J. ; Bonkowsky, Joshua L.
Description:

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.
Subjects:

Animals Article DNA Genotyping Genotyping Techniques High-resolution Melting Analysis (HRMA) Mutation PCR Polymerase Chain Reaction Software Transgenes Zebrafish
Source:

J Vis Exp. 2013; (84):e51138.
Pubmed ID:

24561516
Pubmed Central ID:

PMC4116811
Document Type:

Journal Article
Funding:

DP2 MH100008/DP/NCCDPHP CDC HHS/United States ; DP2 MH100008/MH/NIMH NIH HHS/United States ; R01 MH092256/MH/NIMH NIH HHS/United States
Collection(s):

CDC Public Access
Main Document Checksum:

urn:sha256:1c702d5386bd2af7bfb20d1c445091a287fca9876051f31c53071dea1cbd3566
Download URL:

https://stacks.cdc.gov/view/cdc/30056/cdc_30056_DS1.pdf
File Type:

[PDF - 607.89 KB ]

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