Affinity capture electrophoresis for sequence-specific DNA purification.

A new method, affinity capture electrophoresis (ACE), has been developed for the sequence-specific isolation of DNA. The target DNA is complexed with a biotinylated probe and electrophoresed in a gel equipped with a trap of immobilized streptavidin. This selectively captures the target molecule and its biotinylated probe, while other nontarget molecules pass through the trap. The target DNA is subsequently recovered from the trap by destroying the interaction between the target DNA and the biotinylated probe. Two variations of this technique, one using triple-helix formation and the other using hybridization with a uracil-containing DNA probe at the end of the target fragment, proved effective in model experiments. Since this technique requires no denaturation and handles DNA inside an agarose gel matrix, it is, in principle, applicable to the isolation of very large DNAs.

[1]  S. Strobel,et al.  Site-specific cleavage of a yeast chromosome by oligonucleotide-directed triple-helix formation. , 1990, Science.

[2]  P. Hsieh,et al.  Pairing of homologous DNA sequences by proteins: evidence for three-stranded DNA. , 1990, Genes & development.

[3]  T. Povsic,et al.  Triple helix formation by oligonucleotides on DNA extended to the physiological pH range , 1989 .

[4]  K. Mullis,et al.  Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. , 1987, Methods in enzymology.

[5]  P. Dervan,et al.  Second structural motif for recognition of DNA by oligonucleotide-directed triple-helix formation. , 1991, Science.

[6]  D. Pei,et al.  A combinatorial approach toward DNA recognition , 1991, Science.

[7]  L J Ferrin,et al.  Selective cleavage of human DNA: RecA-assisted restriction endonuclease (RARE) cleavage. , 1991, Science.

[8]  S. J. Flint,et al.  Site-specific oligonucleotide binding represses transcription of the human c-myc gene in vitro. , 1988, Science.

[9]  P. Crosson,et al.  Polyamines favor DNA triplex formation at neutral pH. , 1991, Biochemistry.

[10]  C. Cantor,et al.  A stable complex between homopyrimidine oligomers and the homologous regions of duplex DNAs. , 1988, Nucleic acids research.

[11]  A. Suyama,et al.  A rapid and efficient cloning method with a solid-phase DNA probe: application for cloning the 5'-flanking region of the gene encoding human fibronectin. , 1990, Gene.

[12]  P. Dervan,et al.  Flanking sequence effects within the pyrimidine triple-helix motif characterized by affinity cleaving. , 1992, Biochemistry.

[13]  V. Hořejší Review: Affinity electrophoresis. , 1981, Analytical biochemistry.

[14]  S. Weissman,et al.  Selective enrichment of a large size genomic DNA fragment by affinity capture: an approach for genome mapping. , 1990, Nucleic acids research.