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KMID : 0352519950320020189
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Korea Univercity Medical Journal
1995 Volume.32 No. 2 p.189 ~ p.196
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Construction of Band Specific Painting Probe for Human Chromosome 7q22
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Abstract
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Physical microdissection of metaphase chromosome is the most straightforward approach for the isolation of DNA sequences from specific chromosome regions, it has played a geat role in cloning of genes as well as for cytogenetic analysis of
genetic
diseases and cancers.
In this study we have carried out to construct the band-specific painting probe for 7q22. Microdissection was performed using an inverted microscope (magnification 1000X) with the help of microglass needles and an micromanipulator and we
dissected
25
chromosome fragment from 7q22. Universal amplification of the microdissected fragements with universal primer resulted in a smear ranging from 200-800 base pairs. To prepare painting probe, 2ul of the PCR product were labelled in a secondary PCR
with
biotin 16-dUTP. The probe was hybridized back to normal metaphase chromosomes by fluorescent in situ hybridization and fluorescent signals were detected at 7q22 specifically.
In conclusion, this method will allow the rapid generation of numerous band-specific probes which will faciliate diagnosis of specific genetic diseases, and significantly extends the limits of conventional cytogenetic analysis and thus enabling
the
analysis of unknown chromosome regions.
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KEYWORD
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