PUBLICATION

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

Authors
Xing, L., Quist, T.S., Stevenson, T.J., Dahlem, T.J., Bonkowsky, J.L.
ID
ZDB-PUB-140513-482
Date
2014
Source
Journal of visualized experiments : JoVE   (84): e51138 (Journal)
Registered Authors
Bonkowsky, Joshua, Quist, Tyler
Keywords
none
MeSH Terms
  • Animals
  • DNA/analysis
  • DNA/chemistry
  • DNA/genetics
  • Genotyping Techniques/methods*
  • Polymerase Chain Reaction/methods*
  • Software
  • Zebrafish/genetics*
PubMed
24561516 Full text @ J. Vis. Exp.
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.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping