A dot-blot hybridization method and test kit for detecting normal and mutated genes in unfractionated eucaryotic genomic DNA. Unfractionated single stranded sample DNA is immobilized in a limited zone on a solid support sheet and hybridized with an oligonucleotide probe complementary to the sequence of interest. After stringency washing to remove nonspecifically hybridized probe, the dot-blot zone is examined for the presence of hybridized probe, usually by detection of a label attached to the probe. Hybridization conditions are selected to provide a level of detectable hybridized probe in the dot-blot zone which is readily distinguishable from any background signal associated with the surrounding area on the support sheet. The method eliminates the need for substantial purification and fragmentation of sample DNA required by prior art techniques.
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