Laser microdissection and laser pressure catapulting for the generation of chromosome-specific paint probes.
Chromosome-specific paint probes provide a powerful tool with wide applications in cytogenetic analysis. Here, we present a new approach using UV-laser microbeam microdissection in combination with laser-pressure catapulting, which allows the fast isolation of single chromosomes for the generation of chromosome-specific paint probes. To demonstrate the feasibility of this approach, single chromosomes were collected and amplified with degenerate oligonucleotide-primed PCR, hapten-labeled and hybridized onto normal metaphase spreads. Fluorescence in situ hybridization signals revealed specific painting of the respective chromosomes.
Schermelleh
L
L
Ludwig Maximilians Universität, Munich, Germany.
Thalhammer
S
S
Heckl
W
W
Pösl
H
H
Cremer
T
T
Schütze
K
K
Cremer
M
M
D000818:Animals; D002478:Cells, Cultured; D020223:Chromosome Painting / Q000379:methods*; D006224:Cricetinae; D015342:DNA Probes; D017404:In Situ Hybridization, Fluorescence; D007834:Lasers; D016133:Polymerase Chain Reaction; D014466:Ultraviolet Rays
D015342:DNA Probes