The nematode Caenorhabditis elegans is the First animal whose genome is com
pletely sequenced, providing a rich source of gene information relevant to
metazoan biology and human disease. This abundant sequence information perm
its a broad-based gene inactivation approach in C. elegans, in which chemic
ally mutagenized nematode populations are screened by PCR for deletion muta
tions in a specific targeted gene. By handling mutagenized worm growth, gen
omic DNA templates, PCR screens, and mutant recovery ail in 96-well microti
ter plates, we have scaled up this approach to isolate deletion mutations i
n >100 genes to date. Four chemical mutagens, including ethyl methane sulfo
nate, ethlynitrosourea, diepoxyoctane, and ultraviolet-activated trimethylp
soralen, induced detectable deletions at comparable Frequencies. The deleti
ons averaged similar to 1400 bp in size when using a -3 kb screening window
. The vast majority of detected deletions removed portions of one or more e
xons, likely resulting in loss of gene function. This approach requires onl
y the knowledge of a target gene sequence and a suitable mutagen, and thus
provides a scalable systematic approach to gene inactivation for any organi
sm that can be handled in high density arrays.