Universal probe system for DNA fingerprinting.
A universal probe system is a combination of an unlabeled primary probe that has been inserted in a cloning vector and a labeled secondary probe that is specific to the vector. This system is time- and labor-saving in that one does not need to label probes every time one uses them, so long as they are inserted in the same vector. As a first step in the preparation of a universal probe system for DNA fingerprinting, we isolated multi-locus probes from a subgenomic library that had been constructed by insertion into the phagemid pUC118 of 1-2 kb fragments of DNA from human myeloma cells. Next, we isolated single-stranded DNAs of recombinant phagemids, and hybridized them with Southern blots of DNAs from mother-child-father trios or unrelated individuals. As with double-stranded DNA probes, we were able to detect DNA fingerprints, with a commercially available, alkaline phosphatase-labeled secondary probe or a digoxigenin-labeled universal sequencing primer.
Tamaki
Y
Y
Department of Forensic Medicine, Oita Medical University, Japan.
Kishida
T
T
Fukuda
M
M
D016172:DNA Fingerprinting / Q000379:methods*; D015342:DNA Probes / Q000096:biosynthesis; D015698:Genomic Library; D006801:Humans
D015342:DNA Probes