Mapping individual cosmid DNAs by direct AFM imaging

David P. Allison; Peggy S. Kerper; Mitchel J. Doktycz; Thomas Thundat; Paul Modrich; Frank W. Larimer; Dabney K. Johnson; Peter R. Hoyt; Michael L. Mucenski; Robert J. Warmack

doi:10.1006/geno.1997.4686

Mapping individual cosmid DNAs by direct AFM imaging

David P. Allison, Peggy S. Kerper, Mitchel J. Doktycz, Thomas Thundat, Paul Modrich, Frank W. Larimer, Dabney K. Johnson, Peter R. Hoyt, Michael L. Mucenski, Robert J. Warmack

Sensors and Electronics

Research output: Contribution to journal › Article › peer-review

39 Scopus citations

Abstract

Individual cosmid clones have been restriction mapped by directly imaging, with the atomic force microscope (AFM), a mutant EcoRI endonuclease site-specifically bound to DNA. Images and data are presented that locate six restriction sites, predicted from gel electrophoresis, on a 35-kb cosmid isolated from mouse chromosome 7. Measured distances between endonuclease molecules bound to λ DNA, when compared to known values, demonstrate the accuracy of AFM mapping to better than 1%. These results may be extended to identify other important site-specific protein-DNA interactions, such as transcription factor and mismatch repair enzyme binding, difficult to resolve by current techniques.

Original language	English
Pages (from-to)	379-384
Number of pages	6
Journal	Genomics
Volume	41
Issue number	3
DOIs	https://doi.org/10.1006/geno.1997.4686
State	Published - May 1 1997

Funding

Research at Oak Ridge sponsored by the U.S. Department of Energy by Lockheed Martin Energy Research Corp. under Contract DE-AC05-96OR22464.

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10.1006/geno.1997.4686

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title = "Mapping individual cosmid DNAs by direct AFM imaging",

abstract = "Individual cosmid clones have been restriction mapped by directly imaging, with the atomic force microscope (AFM), a mutant EcoRI endonuclease site-specifically bound to DNA. Images and data are presented that locate six restriction sites, predicted from gel electrophoresis, on a 35-kb cosmid isolated from mouse chromosome 7. Measured distances between endonuclease molecules bound to λ DNA, when compared to known values, demonstrate the accuracy of AFM mapping to better than 1%. These results may be extended to identify other important site-specific protein-DNA interactions, such as transcription factor and mismatch repair enzyme binding, difficult to resolve by current techniques.",

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AU - Allison, David P.

AU - Kerper, Peggy S.

AU - Doktycz, Mitchel J.

AU - Thundat, Thomas

AU - Modrich, Paul

AU - Larimer, Frank W.

AU - Johnson, Dabney K.

AU - Hoyt, Peter R.

AU - Mucenski, Michael L.

AU - Warmack, Robert J.

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AB - Individual cosmid clones have been restriction mapped by directly imaging, with the atomic force microscope (AFM), a mutant EcoRI endonuclease site-specifically bound to DNA. Images and data are presented that locate six restriction sites, predicted from gel electrophoresis, on a 35-kb cosmid isolated from mouse chromosome 7. Measured distances between endonuclease molecules bound to λ DNA, when compared to known values, demonstrate the accuracy of AFM mapping to better than 1%. These results may be extended to identify other important site-specific protein-DNA interactions, such as transcription factor and mismatch repair enzyme binding, difficult to resolve by current techniques.

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Mapping individual cosmid DNAs by direct AFM imaging

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