Process for the preparation of oligonucleotides in solution
First Claim
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1. A process of preparing oligonucleotides in a liquid phase reaction mixture, comprising the steps of:
- a) reacting a bifunctional carrier molecule, which is soluble in, and chromatographically separable from, the reaction mixture, the carrier molecule having the formula;
##STR2## wherein;
(i) n is an integer from 2 to 10; and
(ii) X is a trityl halide;
with at least an equimolar amount of a first nucleoside, nucleotide, or oligonucleotide, N1 ;
b) separating and purifying the compound formed by the reaction in step (a) by chromatography, said compound having the formula;
##STR3## c) repeating steps (a) and (b) to add additional nucleotides N2 . . . Nm to form an oligonucleotide having the formula;
space="preserve" listing-type="equation">N.sup.m . . . N.sup.1 --X--O--C(CH.sub.2).sub.n C--O--X--N.sup.1 . . . N.sup.m wherein m is an integer from 3 to 50 and wherein the oligonucleotide is soluble in and chromatographically separable from the reaction mixture; and
d) releasing the oligonucleotide from the carrier molecule by contacting said oligonucleotide with acetic acid, thereby obtaining an oligonucleotide having the formula;
space="preserve" listing-type="equation">N.sup.1, N.sup.2 . . . N.sup.m.
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Abstract
Process for the preparation of oligonucleotides by successive linking of individual or several nucleoside and/or nucleotide units in a homogeneous phase system using a bi-, tri- or tetra-functional carrier molecule of the general formula Sp(X)n, wherein X is a reactive group which is compatible from the point of view of nucleotide chemistry and n is an integer from 2 to 4, the sequence SP(X-N1 -N2 . . . Nm)n being built up stepwise on the carrier molecule and the oligonucleotide of the formula N1 -N2 . . . Nm subsequently being released.
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8 Claims
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1. A process of preparing oligonucleotides in a liquid phase reaction mixture, comprising the steps of:
- a) reacting a bifunctional carrier molecule, which is soluble in, and chromatographically separable from, the reaction mixture, the carrier molecule having the formula;
##STR2## wherein;
(i) n is an integer from 2 to 10; and(ii) X is a trityl halide; with at least an equimolar amount of a first nucleoside, nucleotide, or oligonucleotide, N1 ; b) separating and purifying the compound formed by the reaction in step (a) by chromatography, said compound having the formula;
##STR3## c) repeating steps (a) and (b) to add additional nucleotides N2 . . . Nm to form an oligonucleotide having the formula;
space="preserve" listing-type="equation">N.sup.m . . . N.sup.1 --X--O--C(CH.sub.2).sub.n C--O--X--N.sup.1 . . . N.sup.mwherein m is an integer from 3 to 50 and wherein the oligonucleotide is soluble in and chromatographically separable from the reaction mixture; and d) releasing the oligonucleotide from the carrier molecule by contacting said oligonucleotide with acetic acid, thereby obtaining an oligonucleotide having the formula;
space="preserve" listing-type="equation">N.sup.1, N.sup.2 . . . N.sup.m. - View Dependent Claims (2, 3)
- a) reacting a bifunctional carrier molecule, which is soluble in, and chromatographically separable from, the reaction mixture, the carrier molecule having the formula;
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4. A process of preparing oligonucleotides in a liquid phase reaction mixture comprising the steps of:
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a) reacting a carrier molecule having the formula;
##STR4## wherein R is --OCH3, with at least an equimolar amount of a nucleoside or nucleotide;b) separating and purifying the compound formed in (a) by gel filtration chromatography c) repeating steps (a) and (b) to form an oligonucleotide which is chromatographably separable from the reaction mixture; and d) releasing the oligonucleotide from the carrier molecule by contacting said oligonucleotide with acetic acid. - View Dependent Claims (5, 6, 7, 8)
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Specification