Generation of Chromosome Painting Probes from Single Chromosomes by Laser Microdissection and Linker-Adaptor PCR

被引：0
作者：
Stefan Thalhammer
Sabine Langer
Michael R. Speicher
Wolfgang M. Heckl
Jochen B. Geigl
机构：
[1] University of Munich,Department für Geowissenschaften
[2] Technische Universität München,Institut für Humangenetik
[3] GSF-Forschungszentrum für Umwelt und Gesundheit,Institut für Humangenetik
来源：
Chromosome Research | 2004年 / 12卷
关键词：
chromosome painting; laser microdissection; linker-adaptor PCR; molecular cytogenetics;
D O I：
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中图分类号：
学科分类号：
摘要：
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.
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页码：337 / 343
页数：6
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