Mapping individual cosmid DNAs by direct AFM imaging.
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 lambda 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.
Allison, DP; Kerper, PS; Doktycz, MJ; Thundat, T; Modrich, P; Larimer, FW; Johnson, DK; Hoyt, PR; Mucenski, ML; Warmack, RJ
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