chromosome walking

A technique that identifies overlapping cloned DNA fragments that form one continuous segment of a chromosome. These fragments can be generated either by random shearing or by partial digestion with a four-base-pair cutter such as Sau3A. A series of colony hybridizations is then carried out, starting with some cloned fragment which has already been identified and which is known to be in the region encompassed by the overlapping clones. This identified fragment is used as a probe to pick out clones containing adjacent sequences. These are then used as probes themselves to identify clones carrying sequences adjacent to them and so on. At each round of hybridization one “walks” further along the chromosome from the initial fragment. See positional cloning.