Selective restriction fragment amplification: fingerprinting

Abstract

The invention relates to a process for the controlled amplification of at least one part of a starting DNA containing a plurality of restriction sites for a determined specific restriction endonuclease, and of which at least part of its nucleic acid is unknown. Application of this process to human, animal or plant DNA fingerprinting, to identification of restriction fragment length polymorphisms. Kit for the application of the process.

1 . A primer, comprising
 (A) a nucleotide sequence that is complementary to at least part of an adaptor sequence;   (B) a nucleotide sequence that is complementary to at least part of a restriction recognition site; and   (C) a selective nucleotide sequence consists of one to 10 nucleotide residues, may be random or specific, and wherein upon hybridization to said target DNA, said selective sequence selects a subset of DNA fragments from the target DNA for amplification or elongation.   
     
     
         2 . A method for identifying an amplified restriction fragment from an individual, comprising:
 (A) digesting DNA obtained from an individual with at least one restriction endonuclease to produce restriction fragments;   (B) ligating a double-stranded adaptor to an end of at least one restriction fragment to produce a tagged restriction fragment; and   (C) amplifying the tagged restriction fragment(s) with two primers at least one of which comprises a sequence complementary to at least part of the adaptor;   wherein the amplification reaction reduces the complexity of the individual's digested DNA and selectively generates a subset of amplified restriction fragments.   
     
     
         3 . The method of  claim 2 , wherein two double-stranded adaptors are ligated to either end of a restriction fragment. 
     
     
         4 . The method of  claim 2 , wherein the individual's DNA is digested with two or more restriction endonucleases. 
     
     
         5 . A method for identifying an amplified restriction fragment from an individual, comprising:
 (A) digesting DNA from an individual with at least one restriction endonuclease to produce restriction fragments;   (B) ligating a double-stranded adaptor to each end of at least one restriction fragment to produce a tagged restriction fragment;   (C) amplifying the tagged restriction fragment(s) with two primers each of which comprises a sequence complementary to at least part of the adaptors   wherein the amplification reaction reduces the complexity of the individual's digested DNA and selectively generates a subset of amplified restriction fragments.