| Construction and Application of Painting Probe for Chromosome Band 1p36.1, 7q11, 7q32-34, 17q12-21. |
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K H Kim, S Y Park, Y H Jeon, S H Park |
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| Abstract |
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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 probes for human chromosomes by microdissection 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 probes for 1p36.1, 7q11, 7q32-34, 17q12-21. The probes has been used successfully to determine the derivation of chromosome segment unidentifiable by standard chromosome banding analysis in IMR-32 cell line and applied to somatic hybrid cell line containing chromosome 1, 7, 17. Thus these painting probes can be applied to detect the related marker chromosomes in many solid tumors, and the band specific probe pools may be used to analyse the genes. |
| Key Words:
microdissection, FISH, IMR-32 cell, Somatic hybrid cell |
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