THE ATOMIC-FORCE MICROSCOPE AS A NEW MICRODISSECTING TOOL FOR THE GENERATION OF GENETIC PROBES

The atomic force microscope (AFM) can be used to visualize and to mani pulate biological material with relative ease and high resolution. Thi s study was carried out to investigate whether probe sets, specific fo r subregions of the human genome and useful for the painting of chromo some bands, can be established by PCR amplification of AFM-dissected c hromosome regions. Compared to standard microdissection techniques, th e AFM can be used with much higher precision for the dissection of the region of interest and subsequent nanoextraction of DNA material. Aft er scanning the area of interest in noncontact mode AFM, chromosome ba nds were cut by the AFM tip at high force. The genetic material of a s ingle cut attached itself to the tip and was extracted and amplified u sing degenerate oligonucleotide-primed-PCR. Subsequent to hapten label ing, fluorescence in situ hybridization was performed and chromosome b and-specific probes mere visualized by standard fluorescence microscop y. (C) 1997 Academic Press.