Production of the F0 Generation by Microinjection of the Gene Trap
6:37
Screening of the F0 Fish
10:00
Screening of the F1 Fish
11:28
Results: Gene Trapping with Gal4-VP16
13:02
Conclusion
This protocol describes the method of gene trap insertional mutagenesis using Gal4-VP16 as the primary reporter and GFP/RFP as secondary reporters in zebrafish. Approximately one in ten high-expressing F0 fish yield gene trap progeny co-expressing GFP and RFP. The screening procedure can be readily scaled to adapt to the size of the laboratory performing the insertional mutagenesis screen.
