Generation of chromosome painting probes from single chromosomes by laser microdissection and linker-adaptor PCR

Chromosome Research : an International Journal on the Molecular, Supramolecular and Evolutionary Aspects of Chromosome Biology
Stefan ThalhammerJochen B Geigl

Abstract

Fluorescence in situ hybridization (FISH) plays an essential role in research and clinical diagnostics. The versatility and resolution of FISH depends critically on the probe set used. Here, we describe an improved approach for the generation of specific DNA probes from single copies of chromosomes. Single chromosomes or single chromosomal regions were microdissected by laser pressure catapulting and amplified using linker-adaptor PCR. The probes were labeled and tested in various scenarios including multicolor-FISH experiments employing up to seven different fluorochromes. FISH confirmed the specific and even staining of the respective chromosomal regions. Furthermore, the capability of these probes to detect even small translocations (<3 Mb) suggests that the dissected regions are completely represented in the generated painting probes.

Citations

Jan 9, 2009·Biomedical Microdevices·Daniela WoideStefan Thalhammer
Apr 5, 2007·Nature Protocols·Jochen B GeiglMichael R Speicher
Jun 29, 2011·Journal of Nanobiotechnology·Sebastiano Di BucchianicoDario Botti
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Jun 25, 2020·Proteomics·Ferdinand von Eggeling, Franziska Hoffmann

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