Abstract
Positional cloning allows the identification of genes through knowledge of their location within the genome. The positional cloning strategy in mice typically involves four distinct steps: (1) the locus is mapped genetically using an intercross or backcross to a genetically divergent strain, (2) DNA probes located close to the locus are obtained, (3) the probes and clone walking is used to identify cloned DNA segments that contain the locus, and (4) genes within the cloned segments are identified and a candidate gene is proposed as the source of the phenotypic change based on expression and functional studies. In mice, positional cloning has had several recent successes in identifying the basis of phenotypes such as iv and lgl , sin, lurcher, and df. This article describes methods used in the positional cloning of df that have general applications, including aspects of intercross development, genetically directed representational difference analysis (GDRDA), candidate gene identification, and the technique of Snorthern analysis, a novel method for examining gene expression in restricted tissue samples.
Published Version
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