|ZFIN ID: ZDB-PUB-140513-482|
Rapid and efficient zebrafish genotyping using PCR with high-resolution melt analysis
Xing, L., Quist, T.S., Stevenson, T.J., Dahlem, T.J., Bonkowsky, J.L.
|Source:||Journal of visualized experiments : JoVE (84): e51138 (Journal)|
|Registered Authors:||Bonkowsky, Joshua, Quist, Tyler|
|PubMed:||24561516 Full text @ J. Vis. Exp.|
Xing, L., Quist, T.S., Stevenson, T.J., Dahlem, T.J., Bonkowsky, J.L. (2014) Rapid and efficient zebrafish genotyping using PCR with high-resolution melt analysis. Journal of visualized experiments : JoVE. (84):e51138.
ABSTRACTZebrafish 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.