METHOD FOR DETERMINING BASE SEQUENCE

摘要

PURPOSE:To simultaneously determine the base sequence of a long DNA by cleaving a specific site of a fluorescently labeled DNA fragment with a restriction enzyme and subsequently subjecting the cleaved products to a gel electrophoresis separation. CONSTITUTION:A specific site of a DNA fragment is cleaved with a restriction enzyme to form shortened DNA fragments, which are subjected to an electrophoresis separation for the determination of the base sequences thereof. The method permits to simultaneously determine the base sequences of long chain DNA fragments having chain lengths of 400-500 which makes the detection difficult or impossible because of the too close electrophoresis patterns of the sequences. The cleavage site of the fluorescently labeled DNA fragment with the restriction enzyme is not specifically limited, but a site giving a base length of 400-500 which is a detection limit by conventional methods is preferable. The restriction enzyme is preferably selected from enzymes having few cleavage sites in 1 to 400 or 500 base length of a specimen DNA which base sequence should be determined.