DNA-DNA hybridization at low temperature using DNA chemically labeled with 14C-dimethyl sulfate |
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| Authors: | H Akiyoshi N Yamamoto |
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| Affiliation: | 1. Fels Research Institute Temple University School of Medicine Philadelphia, Pa. 19140 USA;2. Department of Microbiology Temple University School of Medicine Philadelphia, Pa. 19140 USA;1. EMBL Australia Node in Single Molecule Science, School of Medical Sciences, University of New South Wales, Sydney, Australia;2. ARC Centre of Excellence in Advanced Molecular Imaging, University of New South Wales, Sydney, Australia;1. Department of Biochemistry and Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100005, PR China;2. Yunnan Key Laboratory for The Prevention and Control of Insect-borne Infectious Diseases, Yunnan Malaria Research Center, Yunnan Institute of Parasitic Disease, Pu’er 665000 Yunnan, PR China;3. Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104;5. Department of Biomedical Sciences, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104;4. School of Biomedical Engineering, Drexel University, Philadelphia, Pennsylvania 19104;1. Department of Microbiology and Immunology, Drexel University College of Medicine, Philadelphia, PA, USA;2. Immunotope, Inc., Pennsylvania Institute for Biotechnology, Doylestown, PA, USA;3. Department of Medicine, Imperial College, London, UK;4. Department of Medicine and Department of Laboratory Medicine, University of California at San Francisco, USA;5. Blood Systems Research Institute San Francisco, CA, USA;6. Department of Tropical Medicine, John A. Burns School of Medicine, University of Hawaii, Honolulu, HI, USA |
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| Abstract: | Purified P22 and P221 DNAs were labeled by methylation with 14C-dimethyl sulfate at 4°C. This labeled moiety was unstable at high temperature. About 60% of the radioactivity was released from the DNA by incubation at 65°C for 20 hrs. Using the 14C-methylated DNA, the maximal efficiency of DNA-DNA hybridization was obtained by hybridization at 27°C in the presence of 30% formamide. Determination of the homology between partially genetically related phages proved that this hybridization technique is a useful tool for genetic study. |
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