RT Journal
A1 Liu, Leo X.
A1 Spoerke, Jill M.
A1 Mulligan, Evan L.
A1 Chen, Jing
A1 Reardon, Brian
A1 Westlund, Bethany
A1 Sun, Lin
A1 Abel, Ken
A1 Armstrong, Barbara
A1 Hardiman, Gary
A1 King, Judith
A1 McCague, Lisa
A1 Basson, Michael
A1 Clover, Ralph
A1 Johnson, Carl D.
T1 High-Throughput Isolation of Caenorhabditis elegans Deletion Mutants
JF Genome Research 
JO Genome Research 
YR 1999 
FD September 01 
VO 9 
IS 9 
SP 859 
OP 867 
DO 10.1101/gr.9.9.859 
UL http://genome.cshlp.org/content/9/9/859.abstract 
AB The nematode Caenorhabditis elegans is the first animal whose genome is completely sequenced, providing a rich source of gene information relevant to metazoan biology and human disease. This abundant sequence information permits a broad-based gene inactivation approach in C. elegans, in which chemically mutagenized nematode populations are screened by PCR for deletion mutations in a specific targeted gene. By handling mutagenized worm growth, genomic DNA templates, PCR screens, and mutant recovery all in 96-well microtiter plates, we have scaled up this approach to isolate deletion mutations in >100 genes to date. Four chemical mutagens, including ethyl methane sulfonate, ethlynitrosourea, diepoxyoctane, and ultraviolet-activated trimethylpsoralen, induced detectable deletions at comparable frequencies. The deletions averaged ∼1400 bp in size when using a ∼3 kb screening window. The vast majority of detected deletions removed portions of one or more exons, likely resulting in loss of gene function. This approach requires only the knowledge of a target gene sequence and a suitable mutagen, and thus provides a scalable systematic approach to gene inactivation for any organism that can be handled in high density arrays.