Method of characterizing genomic DNA by reference to a genetic v

Details Method of characterizing genomic DNA by reference to a genetic v Method of characterizing genomic DNA by reference to a genetic v

1992-07-29

1995-05-09

Rollins, John W.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

536 231, 536 242, 536 2431, 935 78, C12Q 168

Patent

active

054139082

ABSTRACT:
It has been a problem that the identification of genomic DNA by restriction fragment length polymorphisms is limited, owing to the low level of genetic variation ordinarily detectable by cloned DNA in this method. The invention provides for improved identification by making use of the existence of DNA regions of hypervariability, otherwise called minisatellite regions in which the DNA contains tandem repeats or quasi-block copolymer sequences and the number of repeats or copolymer units varies considerably from one individual to another. It has now been found that many such regions can be probed simultaneously in such a way as to display this variability using a DNA or other polynucleotide probe of which the essential constituent is a short core sequence tandemly repeated at least 3 and preferably at least 10 times. The probing reveals differences in genomic DNA at multiple highly-polymorphic minisatellite regions to produce an individual-specific DNA "fingerprint" of general use for genetic identification purposes. The "core" used is typically a sequence of 6 to 16 nucleotides contained in or having a high degree of homology with a nucleotide sequence of formula GGAGGTGGGCAGGAXG (2) in which X is A or G, or AGAGGTGGGCAGGTGG (3) or GGAGGYGGGCAGCAGG (4) in which Y is C or T, or with a 12-15 nucleotide sequence of formula T(C).sub.m GGAGGAXGG(G).sub.p C (5A) or T(C).sub.m GGAGGA(A).sub.q GGGC (5B) in which X is A or G, M is 0, 1 or 2, p is 0 or 1, and q is 0 or 1, or GPGGGCWGGWXG (6) in which X is as above, P is not G, and W=A or G. The invention is particularly useful in paternity and maternity testing, forensic medicine and the diagnosis of genetic diseases and cancer.

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