Generation of chromosome painting probes from single chromosomes by laser microdissection and linker-adaptor PCR.
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.
Thalhammer
Stefan
S
Department für Geowissenschaften, University of Munich, Theresienstr. 41, D-80333 Munich, Germany. s.thalhammer@lrz.uni-muenchen.de
Langer
Sabine
S
Speicher
Michael R
MR
Heckl
Wolfgang M
WM
Geigl
Jochen B
JB
D020223:Chromosome Painting / Q000379:methods; D002875:Chromosomes / Q000235:genetics*; D015342:DNA Probes / Q000235:genetics*; D005456:Fluorescent Dyes; D006801:Humans; D042282:Microdissection; D016133:Polymerase Chain Reaction; D014178:Translocation, Genetic / Q000235:genetics
D015342:DNA Probes; D005456:Fluorescent Dyes