A method for double-stranded DNA purification, by which a solution containing DNA in a mixture with other components is passed over a support on which is covalently coupled an oligonucleotide capable of hybridizing with a specific sequence present on the DNA to form a triple helix.
대표청구항▼
1. A triple helix bound to a purification support that is a functionalized chromatographic support, wherein the triple helix comprises a pyrimidine-rich oligonucleotide covalently coupled directly to the support through an amide or amine link and a double-stranded DNA that has hybridized to the olig
1. A triple helix bound to a purification support that is a functionalized chromatographic support, wherein the triple helix comprises a pyrimidine-rich oligonucleotide covalently coupled directly to the support through an amide or amine link and a double-stranded DNA that has hybridized to the oligonucleotide. 2. The triple helix according to claim 1, wherein the oligonucleotide comprises a cytosine-rich sequence. 3. The triple helix according to claim 1, wherein the oligonucleotide comprises a thymidine-rich sequence. 4. The triple helix according to claim 1, wherein the oligonucleotide comprises either a first sequence (CCT)n, a second sequence (CT)n, a third sequence (CTT)n, or a combination of any of the first, second, and third sequences, wherein n is an integer between 1 and 15 inclusive. 5. The triple helix according to claim 1, wherein the support further comprises an arm comprising a carbon chain (CH2)n, wherein n is an integer between 1 and 18 inclusive, and wherein the arm is linked to the oligonucleotide through a phosphate and to the support through an amide link. 6. The triple helix according to claim 5, wherein n is an integer between 6 and 12. 7. A method of making a support for purifying double-stranded DNA, comprising covalently coupling an oligonucleotide to a functionalized chromatographic support, wherein the oligonucleotide comprises a pyrimidine-rich sequence, and forming a triple helix with a double-stranded DNA by hybridizing the oligonucleotide with a nucleotide sequence in the double-stranded DNA, and wherein the oligonucleotide is covalently coupled directly to the support through a disulphide, thioether, ester, amide or amine link. 8. The method according to claim 7, wherein the oligonucleotide comprises a cytosine-rich sequence. 9. The method according to claim 7, wherein the oligonucleotide comprises a thymidine-rich sequence. 10. The method according to claim 7, wherein the oligonucleotide comprises either a first sequence (CCT)n, a second sequence (CT)n, a third sequence (CTT)n, or a combination of any of the first, second, and third sequences, wherein n is an integer between 1 and 15 inclusive. 11. The method according to claim 7, wherein the oligonucleotide is coupled to the support through an amide or amine link. 12. The method according to claim 7, wherein the oligonucleotide is coupled to the support via an arm comprising a carbon chain (CH2)n, wherein n is an integer between 1 and 18 inclusive, and wherein the arm is linked to the oligonucleotide through a phosphate and to the support through an amide link. 13. The method according to claim 12, wherein n is an integer between 6 and 12. 14. The support obtained by the method of claim 7. 15. A method of coupling an oligonucleotide to a purification support, wherein the support is a functionalized chromatographic support that comprises residues, comprising: a) activating the residues of the support, andb) contacting the activated residues of the support with the oligonucleotide to obtain covalent coupling directly between the oligonucleotide and the residues of the purification support; wherein the oligonucleotide comprises a pyrimidine-rich sequence which forms a triple helix with a double-stranded DNA by hybridizing with a nucleotide sequence in the double-stranded DNA. 16. A functionalized chromatographic support comprising a covalently coupled oligonucleotide, wherein the oligonucleotide comprises a pyrimidine-rich sequence which forms a triple helix with a double-stranded DNA by hybridizing with a nucleotide sequence in the double-stranded DNA, wherein the oligonucleotide is covalently coupled directly to the purification support through a disulphide link.
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이 특허에 인용된 특허 (19)
Soubrier,Fabienne, Circular DNA molecule having a conditional origin of replication, process for their preparation and their use in gene therapy.
Crouzet, Jo?l; Soubrier, Fabienne, Circular DNA molecule with conditional origin of replication, method for preparing the same and use thereof in gene therapy.
Cantor Charles R. (Boston MA) Ito Takashi (Chiba MA JPX) Smith Cassandra L. (Boston MA), DNA purification by triplex-affinity capture and affinity capture electrophoresis.
Ghosh Soumitra S. (San Diego CA) Fahy Eoin D. (San Diego CA), End-attachment of oligonucleotides to polyacrylamide solid supports for capture and detection of nucleic acids.
Miller Paul S. (Baltimore MD) Bhan Purshotam (Baltimore MD), Formation of triple helix complexes for the detection of double stranded DNA sequences using oligomers which comprise an.
Froehler Brian C. (Belmont CA) Kent Kenneth M. (Mt. View CA) Wu Sylvia (Castro Valley CA), Large scale synthesis of oligonucleotides and their associated analogs.
Letsinger Robert L. (Wilmette IL) Gryaznov Sergei M. (San Mateo CA), Oligodeoxyribonucleotides including 3′-aminonucleoside-phosphoramidate linkages and terminal 3′-amino groups.
Crouzet,Jo챘l; Scherman,Daniel; Wils,Pierre; Blanche,Fran챌is; Cameron,B챕atrice, Purification of a triple heli formation with an immobilized oligonucleotide.
Crouzet Joel,FRX ; Scherman Daniel,FRX ; Wils Pierre ; Blanche Fran.cedilla.is,FRX ; Cameron Beatrice,FRX, Purification of a triple helix formation with an immobilized oligonucleotide.
Crouzet, Joël; Scherman, Daniel; Wils, Pierre; Blanche, Françis; Cameron, Béatrice, Purification of a triple helix formation with an immobilized oligonucleotide.
Ji Huamin (4817 Sheboygan Ave. Madison WI 53705) Smith Lloyd M. (1115 Amherst Dr. Madison WI 53705), Rapid purification of circular DNA by triplex-mediated affinity capture.
Jendrisak Jerome J. (Madison WI) Lewis Martin K. (Madison WI) Fiandt Michael J. (Cambridge WI), Vectors for in vitro production of RNA copies of either strand of a cloned DNA sequence.
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