Forward Genetic Screening of Insertional Mutants

Insertional mutagenesis has contributed to the success of forward genetics in Arabidopsis thaliana. The availability of large collections of lines mutagenized by either transposon or T-DNA insertions, in combination with the systematic sequencing of insertion/genome junctions, enables the identification of mutations for any given gene in the nuclear genome of this plant species. Protocols for the identification and confirmation of mutations by forward genetics, the isolation of insertion/genome junctions, and the stabilization of transposon-induced mutations are provided, together with an overview of available insertional mutant collections and of their characteristics.