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Two-Dimensional Agarose Gel Electrophoresis as a Tool to Isolate Genus- and Species-Specific Repetitive DNA Sequences

✍ Scribed by Carsten Harms; Ingo Klarholz; Armin Hildebrandt


Publisher
Elsevier Science
Year
2000
Tongue
English
Weight
712 KB
Volume
284
Category
Article
ISSN
0003-2697

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✦ Synopsis


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.