C. elegans deletion mutant screening.
The methods used by the Caenorhabditis elegans Gene Knockout Consortium are conceptually simple. One does a chemical mutagenesis of wild-type C. elegans, and then screens the progeny of the mutagenized animals, in small mixed groups, using polymerase chain reaction (PCR) to identify populations with animals where a portion of DNA bounded by the PCR primers has been deleted. Animals from such populations are then selected and grown clonally to recover a pure genetic strain. We categorize the steps needed to do this as follows: (1) mutagenesis and DNA template preparation, (2) PCR detection of deletions, (3) sibling selection, and (4) deletion stabilization. These are discussed in detail in this chapter.
Barstead
Robert J
RJ
Oklahoma Medical Research Foundation, Oklahoma City, USA.
Moerman
Donald G
DG
D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics; D001483:Base Sequence; D017173:Caenorhabditis elegans / Q000235:genetics*; D004252:DNA Mutational Analysis; D017238:Genes, Helminth / Q000235:genetics; D016296:Mutagenesis; D017384:Sequence Deletion