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

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:
  • Article
  • genotyping
  • high-resolution melting analysis (HRMA)
  • PCR
  • zebrafish
  • mutation
  • transgenes
  • Animals
  • DNA
  • Genotyping Techniques
  More +
• 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
• Download URL:
  https://stacks.cdc.gov/view/cdc/30056/cdc_30056_DS1.pdf
• File Type:
  [PDF - 607.89 KB]
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:1c702d5386bd2af7bfb20d1c445091a287fca9876051f31c53071dea1cbd3566