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Dissection and high-yield recovery of nanometre-scale chromosome fragments using an atomic-force microscope

K Tsukamoto1, S Kuwazaki2, K Yamamoto2, T Ohtani1 and S Sugiyama1

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We developed a new method to dissect and recover nanometre-scale chromosome fragments from specific regions of a chromosome by atomic-force microscopy (AFM) with a novel AFM cantilever control. Recovery of the chromosome fragments using this method was highly reproducible, with a success rate greater than 90%. The width of the dissection depended on the cantilever-tip diameter; similar widths (240 ± 92 nm on average) were obtained when a standard silicon cantilever was used. We also achieved successive dissection and recovery of five fragments located on a specific region of a chromosome. Rinsing the specimen with a biological buffer, such as PBS, facilitated the recovery. This treatment may cause the chromosome structures to be more relaxed. The method, called the 'scratch method', will be a useful tool to easily obtain nucleotide sequences with positional information on a specific region of a chromosome and will contribute to various fields of genetic investigations.


PACS

87.16.Sr Chromosomes, histones

87.64.Dz Scanning tunneling and atomic force microscopy

87.14.G- Nucleic acids

Subjects

Medical physics

Biological physics

Dates

Issue 5 (14 March 2006)

Received 21 September 2005, in final form 9 November 2005

Published 10 February 2006



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