Electron microscopic visualization of restriction sites on DNA molecules. |
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Authors: | T Miwa M Takanami H Yamagishi |
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Institution: | 1. Department of Biophysics, Faculty of Science, Kyoto University, Kyoto, Kyoto 606 Japan;2. Institute for Chemical Research, Kyoto University, Uji, Kyoto 611 Japan |
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Abstract: | DNA molecules were adsorbed to a polylysine-treated carbon film and digested directly on the film by restriction enzymes. After washing the film with 1 M NaCl, 0.4% Kodak Photo-Flo and 9% formamide, each cleavage site introduced was visualized as a gap under the electron microscope. By measuring the gapped positions on linear DNA molecules induced by other enzymes, a single EcoRI site on a lambda dv1 molecule and three HinHI sites on an fd1RF molecule were mapped at the positions expected from the cleavage maps, respectively. This electron-microscopic procedure may be useful for the construction of a cleavage map. |
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Keywords: | Polylysine adsorption Λdv1 and fd in situ digestion DNA gap at cleavage sites |
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