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Genomic subtractive hybridization to isolate species‐specific DNA sequences in insects
Author(s) -
Clapp J. P.,
McKee R. A.,
AllenWilliams L.,
Hopley J. G.,
Slater R. J.
Publication year - 1993
Publication title -
insect molecular biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.955
H-Index - 93
eISSN - 1365-2583
pISSN - 0962-1075
DOI - 10.1111/j.1365-2583.1993.tb00114.x
Subject(s) - biology , genomic dna , dna , suppression subtractive hybridization , biotinylation , genetics , homologous chromosome , dna sequencing , melanogaster , dna–dna hybridization , genomic library , microbiology and biotechnology , complementary dna , drosophila melanogaster , gene , base sequence , cdna library
Selective enrichment has been used in a number of instances for the isolation of species‐specific sequences in prokaryotes. This paper reports the successful application of the technique to insects. Genomic probes were derived to the target species D. funebris and D. simulans . The method involves the biotinylation of non‐target ‘driver’ DNA prepared from the closely related species D. melanogaster and its hybridization to homologous sequences in the target DNA. Hybrid molecules were removed from the reaction by incubation with streptavidin followed by phenol extraction, leaving a preparation enriched for target fragments. All DNA fragments isolated in the D. funebris experiments proved to be specific to that species. Five out of twenty‐four fragments screened in the D. simulans experiments were specific when screened with homologous DNA and genomic DNA from its sibling species, D. melanogaster .