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 manipulate biological material with relative case and high resolution. This study was carried out to investigate whether probe sets, specific for subregions of the human genome and useful for the painting of chromosome bands, can be established by PCR amplification of AFM-dissected chromosome regions. Compared to standard microdissection techniques, the AFM can be used with much higher precision for the dissection of the region of interest and subsequent nanoextraction of DNA material. After scanning the area of interest in noncontact mode AFM, chromosome bands were cut by the AFM tip at high force. The genetic material of a single cut attached itself to the tip and was extracted and amplified using degenerate oligonucleotide-primed-PCR. Subsequent to hapten labeling, fluorescence in situ hybridization was performed and chromosome band-specific probes were visualized by standard fluorescence microscopy.
Thalhammer
S
S
Institut Für Kristallographie und Angewandte Mineralogie, Universität München; Germany.
Stark
R W
RW
Müller
S
S
Wienberg
J
J
Heckl
W M
WM
D002889:Chromosomes, Human, Pair 2 / Q000737:chemistry / Q000648:ultrastructure*; D004247:DNA; D004249:DNA Damage; D015342:DNA Probes; D005456:Fluorescent Dyes; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D018625:Microscopy, Atomic Force / Q000379:methods; D008856:Microscopy, Fluorescence; D016133:Polymerase Chain Reaction
D015342:DNA Probes; D005456:Fluorescent Dyes; D004247:DNA