Two-dimensional agarose gel electrophoresis as a tool to isolate genus- and species-specific repetitive DNA sequences.
Two-dimensional electrophoresis in agarose gels separates DNA-restriction fragments not only by molecular weight but also according to their AT-cluster content. The method produced genus-specific spot patterns of multicopy DNA fragments of grains as well as spot patterns of highly repetitive DNA fragments of ciliates, demonstrated for barley, spelt, and Tetrahymena. Further investigations in regard to their specificity by hybridization with three other grain species (wheat, oat, and rye) and three ciliate species (Tetrahymena thermophila, Tetrahymena pigmentosa, and Tetrahymena borealis) were performed. The DNA samples from spelt and Tetrahymena were demonstrated to be genus specific for Triticum and species specific for Tetrahymena pyriformis, respectively.
University of Bremen, FB2, UFT, Room 1100, Bremen, 28359, Germany. carsten@uni-bremen.de
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Harms
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Klarholz
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Hildebrandt
D000818:Animals; D018554:Avena / Q000235:genetics; D015139:Blotting, Southern; D018744:DNA, Plant / Q000235:genetics; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D004996:Ethidium / Q000494:pharmacology; D005456:Fluorescent Dyes / Q000494:pharmacology; D001467:Hordeum / Q000235:genetics; D009693:Nucleic Acid Hybridization; D012091:Repetitive Sequences, Nucleic Acid; D012434:Secale / Q000235:genetics; D012680:Sensitivity and Specificity; D013768:Tetrahymena / Q000235:genetics; D016808:Tetrahymena thermophila / Q000235:genetics; D014908:Triticum / Q000235:genetics
D018744:DNA, Plant; D005456:Fluorescent Dyes; D004996:Ethidium