| Construction and Application of Painting probe for
Chromosome band 5q14-q15, 6q23, 16p11.2 and 22q12 |
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Soo-Yeun Park1, So Young Park1, Joon-Il Kil1, Yong-Hyuck Chun1, Sun-Hwa Park2 |
1Institute of Human Genetics, Department of Anatomy, College of Medicine, Korea University, Seoul, Korea
2Department of Anatomy, College of Medicine, Korea University, Seoul, Korea |
| 염색체띠 5q14-q15, 6q23, 16p11.2 및 22q12에 대한 페인팅 probe 제작과 적용 |
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박수연1, 박소영1, 길준일1, 전용혁1, 박선화2 |
1고려대학교 의과대학 해부학교실, 유전병연구소
2고려대학교 의과대학 해부학교실 |
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| Abstract |
Purpose
: Chromosome microdissection has become a very powerful approach to generate chromosome band-specific library and painting probes for physical mapping or cytogenetic analysis. We have constructed here band-specific painting probes for human chromosomes by microdissection and polymerase chain reaction(PCR).
Methods
: We pretreated the microdissected fragments with Topoisomerase I(Topo I) which catalyzes the relaxation of supercoiled DNA, and performed two initial rounds of DNA synthesis with T7 DNA polymerase followed by conventional PCR to enable the reliable preparation of fluorescent in situ hybridization(FISH) probe.
Results
: It was possible to construct region-specific painting probes for 5q14-q15, 6q23, 16p11.2 and 22q12. The painting probes were successful in determining the corresponding chromosome bands in SNU-484 cell line, IMR-32 cell line.
Conclusion
: These painting probes can be used to detect the related marker chromosomes in congenital malformations and other solid tumors, and the band specific probe pools may be used effectively to analyse the genes. |
| Key Words:
Chromosome microdissection, PCR, Band-specific painting probe, FISH |
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