PCR-Based Identification of T-DNA Insertion Mutants

With large-scale sequencing projects (1 ,2 ), thousands of unique sequences ESTs; expressed sequence tags) have been generated. In addition, hundreds of sequences have been identified based on then homology with a gene of known interest or from subtractive hybridization procedures. For the vast majority of these genes, the sequence indicates little or nothing about its function in the plant. In thus protocol, we describe a reverse genetics procedure which will be useful for identifying an insertion mutant in Arabidopsis for any sequence of interest.