Korean J Anat.  1998 Oct;31(5):753-760.

Construction of Painting probe for Chromosome Band 18p11.1-p11.2

• Seo YS
• Chun YH
• Park SY
• Kim H
• Park SH

Abstract

Chromosome microdissection has become a very powerful approach to generate chromosome band-specific library and painting probes for physical mapping or cytogenetic analysis. Here we have constructed the band-specific painting probe for human chromosome 18p11.1-p11.2 by microdissec-tion and polymerase chain reaction (PCR). 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 from a single microdissected chromosome. With this method, it was possible to construct region-specific painting probe for 18p11.1-p11.2. The probe has been used successfully to determine the trisomy 18. Thus this painting probe can be applied to detect the related marker chromosomes in various solid tumors, and the band specific probe pools may be used to analyse the genes.

Keyword

chromosome microdissection; 18p11.1-p11.2; fluorescent in situ hybridization; trisomy 18

MeSH Terms

Chromosomes, Human
Cytogenetic Analysis
DNA
DNA Topoisomerases, Type I
DNA, Superhelical
Humans
In Situ Hybridization, Fluorescence
Microdissection
Paint*
Paintings*
Polymerase Chain Reaction
Relaxation
Trisomy
DNA
DNA Topoisomerases, Type I
DNA, Superhelical