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Generation of FISH probes using Laser Microbeam Microdissection and application to clinical molecular cytogenetics

Title
Generation of FISH probes using Laser Microbeam Microdissection and application to clinical molecular cytogenetics
Author
이철훈
Keywords
Chromosome painting; fluorescence in situ hybridization; Laser microbeam microdissection; marker chromosome; polymerase chain reaction
Issue Date
2007-07
Publisher
KOREAN SOC MICROBIOLOGY & BIOTECHNOLOGY
Citation
JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY, v. 17, No. 7, Page. 1079-1082
Abstract
Chromosome microdissection and the reverse FISH technique is one of the most useful methods for the identification of structurally abnormal chromosomes. In particular, the laser microbeam microdissection (LMM) method allows rapid isolation of a target chromosome or a specific region of chromosomes without damage of genetic materials and contamination. Isolated chromosomes were directly amplified by the degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR), and then the FISH probes labeled with spectrum green- or spectrum red-dUTP were generated by nick-translation. Whole chromosome painting (WCP) probes were successfully generated from only 5 copies of the chromosome. With this method, we produced 24 WCP probes for each human chromosome. We also tried to characterize a marker chromosome, which seemed to be originated from chromosome 11 on conventional banding technique. The marker chromosomes were isolated by the LMM method and analyzed by reverse FISH. We elucidated that the marker chromosome was originated from the short ann of chromosome 5 (5p 11 -> pter). A fully automated and computer-controlled LMM method is a very simple laboratory procedure, and enables rapid and precise characterization of various chromosome abnormalities.
URI
http://kiss.kstudy.com/thesis/thesis-view.asp?key=2627691http://repository.hanyang.ac.kr/handle/20.500.11754/106696
ISSN
1017-7825; 1738-8872
Appears in Collections:
COLLEGE OF PHARMACY[E](약학대학) > PHARMACY(약학과) > Articles
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