Characterization Of Microdissected Human Chromosome Region 3p14-24

Microdissection of chromosoies is a recently developed method to provide chromosome region specific DNA libraries which are very useful in genome analysis including physical mapping of unknown genes responsible for different genetic diseases. It was firstly applied to microcloning specific region of Drosophila genome, and then of mammalian and human metaphase chroiosoies. PCR (polymerase chain reaction) which may augment a single copy of DNA template to a huge number was introduced to amplify the microdissected DNA, and thus made this method more applicable to different areas of molecular genetics.Ten to thirty pieces of human chromosome region 3pl4-24 were microdissected and pooled to be digested enzymatically. The obtained DNA were amplified by PCR using a single primer. The PCR products in bulk hybridized readily to human chromosome 3p with FISH(fluorescent in situ hybridization)technique. About 10~4 colonies of a genomic DNA library were screened with these PCR products, 65 clones were positively hybridized. The inserts of 10 clones were analysed with Bam HI digestion, and three of them contained internal Bam HI digestion sites. Two of them (2/3) showed positive results in FISH with human chromosome 3p, and the third one failed to hybridize with chromosome 3p owing to technical accident.