All issues > Volume 43(2); 2000

Original Article

J Korean Pediatr Soc. 2000;43(2):179-187. Published online February 15, 2000.

Construction and Application of Painting probe for Chromosome band 5q14-q15, 6q23, 16p11.2 and 22q12

Soo-Yeun SY Park¹, So Young SY Park¹, Joon-Il JI Kil¹, Yong-Hyuck YH Chun¹, Sun-Hwa SH 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

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.

Keywords :Chromosome microdissection, PCR, Band-specific painting probe, FISH