DE177497T1 - IMMOBILIZATION OF NUCLEIC ACIDS. - Google Patents

Claims (5)

1. nuki einsäure-Matriχ syst em comprising: a solid ^support:; a coupler including a DNA molecule with a first and second ends, the first end being covalently bonded to the solid support and the second End is not bound to the solid support and is a limiting enzyme half-site; and a DNA molecule with a first end to that second end of the coupler complementary delimitation half-point, with the molecule at the complementary half-position is coupled to the second end of the coupler. 2. Matrix system according to claim 1, wherein the coupler is one from naturally occurring DNA by successive Treatment of the DNA with two different limiting endonucleases isolated boundary fragment includes. 3. Matriχ sy st em according to claim 2, wherein the course occurring DNA obtained from the DNA of the plasmid pl) C9 has been. 4. The matrix system of claim 3, wherein the two ^ various limiting endonucleases BgI I and Hind III are. 5. The matrix system of claim 1, wherein the coupler is oligonucleotide and mononucleotide, wherein the .- 'Wo.nonucleotide in vitro chemically bound to the oligonucleotide is fwörden. 6. Matrix system according to claim 5, wherein the oligonucleotide a synthetic egg rvz'ei strand with the base sequence 5'-d (fpCp & pG) -3 * is. ■■? ■. Matrix system according to claim 6, wherein the oligonucleotide at his 3'-OH end a bi ot i nyL i ertes Ribonuk Leot i d has been oathed. so. The matrix system of claim 7, wherein the solid support Agarose is bound to the Streptavidiη kovaLent and wherein the biotinyLierte RibonukIeotid by condensation uh that St reptavid i η has been tied. 9. Matrix system according to claim 5, wherein the OLigonukLeotid a ribonucleotide bound at its 3 * -0H end having. 10 .Mat r i χ sys t em according to claim 9, .With the r solid support ZeLLüLose is covaLent bound to the adipic acid hydrazide is and where the Ribonuk leot id by perj odatox idation has been converted into a D.ialdehyde and the diaidehyde being converted to the adipic acid hydrazide by condensation : has been bound. 11. The matrix system of claim 1, wherein the solid support Cellulose is covalently bound to the adipic hydrazide is. O Ί 77497 12. The matrix system of claim 1, wherein the coupler is a Boundary fragment and a mononucleotide comprises / wherein the mononucleotide has been chemically bound to the delimitation fragment in vitro. 13. Matrix system according to claim 12, wherein the delimitation fragment has a ^{ribonucleotide bound to its 3%} OH end. 14. Matrix system according to claim 13, wherein the solid carrier is cellulose, other Adi pi nsäu rehydrazid is covalently bound, wherein the RibonukIeotid has been converted by periodate oxidation in a dialdehyde and wherein the dialdehyde has been bound by condensation to the adipic acid hydrazide. 15. Matrix system according to claim Λ, wherein the solid support is agarose / is covalently bound to the streptavidin. 16. Coupler matrix system comprising: a solid support; and a coupler having one end covalently bonded to the solid support and a portion which extends from the linked end and contains a limiting enzyme recognition post; wherein the Rekognitionsstel Ie can be activated to convert the Rekogni ti onsstel Ie into a limiting enzyme half-position. 17. The matrix system of claim 16, wherein the coupler comprises a delimitation fragment isolated from naturally occurring DNA by sequentially treating the DNA with two different delimitation endonucleases. -A- 18. Matrix system according to claim 16, wherein the coupler comprises a limiting fragment and a mononucleotide, wherein the mononucleotide is chemically linked to the limiting fragment in vitro _; has been. The matrix system of claim 16, wherein the solid support Cellulose is bound to the adipinic acid rehydrazi d kpyaL ent is. 20th _; Matrix system according to claim 17, wherein the naturally arriving DNA has been obtained from the DNA of the plasmid pUC.9. 21. The matrix system of claim 20, wherein the two various limiting endonucleases BgI I and Hind III are. 22> Matrix system according to claim 18, wherein, the boundary fragment a ribonucleptide is added to its 3'-OH end has been. 23 :. ; Mat ri χ system according to claim 22, wherein the solid support is cellular, on which adipic acid hydrazide is covalently bonded, where the ri bonuk I eot id by periodato, xidate .. i.on has been converted into a dialdehyde and the dialdehyde has been bonded to the adipic acid hydrazide by condensation. 2.4. Kup.pl e r-Nuk Lei nsäu re-Sy st em, comprising: a DNA MoLeküI with a first end with a ... limitation of the enzyme holding point; and a coupler including a second DNA molecule having a first end with one to the delimiting half-position of the first DNA molecule complementary Limiting enzyme halves, with the first end is coupled to the end of the first DNA molecule / and a second end connectable to a solid support. 2 5. Nucleic acid system according to claim 24, wherein the coupler an OLigonucleotide to which biotin has been added is. 26. The nucleic acid system of claim 24, wherein the DNA is a limiting endonuclease fragment. 27. Nucleic acid system according to claim 25, wherein the oligonucleotide is a single strand with the base sequence 5 ^% -d (TpCpGpG) -3 ^l . 28. The nucleic acid system of claim 26, wherein the limiting endonucL ease Aval is. 29. Method of Tying an Exposed Half-Piece a delimitation point for a given delimitation endonuclease to a solid support comprising, in any Order that covalent binding of a coupler, including a OLigonucleotide, which contains the delimitation point, to the solid support; and Splitting of the oligonucleotide at the delimitation point with the limiting endonuclease to remove the exposed To generate half places. 30. The method of claim 29, wherein the solid support a matrix of cellulose or agarose adipic acid dehydrazide and chemical bonding of the BgI I end of the boundary fragment to the solid support w i r d through Addition of a ribonucleus to the end of the BgI I half-position of the delimiting fragment; Oxidation of the ribonucleotide with periodate to reduce it Converting ribose-cis-di-oil into a dialdehyde; and Mixing the boundary ■ fragment with the matrix. 31. Method of Tying an Exposed Half-El Ie a restriction site for each of the desired restriction endonucleases Hind III, Pst I, H inc II, BamH I, Sma I or EcoR I to a solid support comprising isolating a Bgl I / Hind III boundary fragment from plasmid pUC9 DNA; chemically bonding the BgI I end of the limiting fragment to the solid support; and Cleaving the boundary fragment with the boundary endonuclease for which an exposed half- digit is desired. 32. A method of immobilizing DNA on a solid support, comprising the stable binding to the solid support via one end of an oligonucleotide which at its other end has a first half of a known delimitation endonuclease and for which the DNA contains a recognition site for the delimitation endonuclease; Columns of the boundary point on the DNA to the limit endonuclease to form a second and a third half-site on the DNA; and Connect the first half with either the second or the third half by treatment with DNA gas. 33. Method of immobilizing DNA on a solid Carrier, including that Binding the DNA to an oligonucleotide coupler corresponding delimitation endonuclease halves, located at one end of the DNA and a first end of the coupler; and stable · binding of the coupler to the solid support over a second end of the coupler.