C. Elegans Deletion Mutant Screening

Methods Mol Biol. 2006;351:51-8. doi: 10.1385/1-59745-151-7:51.

Abstract

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.

Publication types

  • Review

MeSH terms

  • Animals
  • Animals, Genetically Modified / genetics
  • Base Sequence*
  • Caenorhabditis elegans / genetics*
  • DNA Mutational Analysis
  • Genes, Helminth / genetics
  • Mutagenesis*
  • Sequence Deletion*