Two-dimensional gel electrophoretograms of human chromosome specific restriction DNA fragments.
Human genomic DNA can be scanned by a two-dimensional gel electrophoresis of NotI cleaved, end-labelled DNA. This method, called Restriction Landmark Genome Scanning (RLGS) method yields about 2000 landmark spots in such a two-dimensional gel electrophoresis. To assign these spots to chromosomes, DNA from flow-sorted chromosome 3 (7.7 x 10(5) copies) and chromosome 4 (6.7 x 10(5) copies) were subjected to the same analysis. Ninety seven and 122 spots, respectively, were obtained in the electrophoretograms among which 16 spots were common to other chromosomes, and the others were unique to each chromosome. A method has been devised to assign these spots to the whole genomic profile. The ability to assign each spot to a chromosome will render this new mapping technology extremely useful, in which one can detect physical aberrations occurring in chromosomes, or it can be used as an auxiliary method in physical mapping of the human genome.
Yoshikawa
H
H
Institute for Molecular and Cellular Biology, Osaka University, Japan.
Nagai
H
H
Matsubara
K
K
Fujiyama
A
A
D001345:Autoradiography / Q000379:methods; D002461:Cell Line, Transformed; D002877:Chromosomes, Human / Q000737:chemistry / Q000648:ultrastructure*; D004247:DNA / Q000737:chemistry* / Q000302:isolation & purification; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D006801:Humans; D008297:Male; D015183:Restriction Mapping; D014316:Tritium
D014316:Tritium; D004247:DNA; D015252:Deoxyribonucleases, Type II Site-Specific; C038565:GCGGCCGC-specific type II deoxyribonucleases