Overview
Our protocol for generating deletion mutations is based on the procedures of Bob Barstead and Gary Moulder at the Oklahoma Medical Research Foundation in Oklahoma City. The procedure is to mutagenize a large population of worms with trimethylpsoralen and UV irradiation, set up 1152 subpopulations, screen DNA made from this library for deletions in specific genes by nested PCR, and then to recover single worms carrying the deletions through a sib-selection process. We have made various modifications to the original protocol, specifically with regard to library complexity (100,000 genomes versus 500,000 to 1,000,000) and screening strategy (the use of poison primers to increase sensitivity of detection for small deletions).
Details
Details of this protocol, C. Elegans Gene Knockout Protocol, are located on a web site other than Biocompare Protocols.