Linkers and co-coupling agents for optimization of oligonucleotide synthesis and purification on solid supports
First Claim
1. A method of generating purified oligonucleotides comprising:
- a) providing;
i) a substrate comprising an array of oligonucleotides attached to said substrate via anchor moieties attached to non-cleavable linkers attached to said substrate, wherein said anchor moieties comprise the structure —
C(X)—
C(Y), wherein X comprises —
OPO2O—
, Y is a nucleophile and wherein said structure is part of a ring moiety;
ii) a deprotecting solution; and
iii) and a wash solution;
b) deprotecting said oligonucleotides with said deprotecting solution;
c) washing said oligonucleotides with said wash solution; and
d) cleaving said oligonucleotides at said anchor group to provide purified oligonucleotides.
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Abstract
A method of modulation of synthesis capacity on and cleavage properties of synthetic oligomers from solid support is described. The method utilizes linker molecules attached to a solid surface and co-coupling agents that have similar reactivities to the coupling compounds with the surface functional groups. The preferred linker molecules provide an increased density of polymers and more resistance to cleavage from the support surface. The method is particularly useful for synthesis of oligonucleotides, oligonucleotides microarrays, peptides, and peptide microarrays. The stable linkers are also coupled to anchor molecules for synthesis of DNA oligonucleotides using on support purification, eliminating time-consuming chromatography and metal cation presence. Oligonucleotides thus obtained can be directly used for mass analysis, DNA amplification and ligation, hybridization, and many other applications.
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Citations
19 Claims
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1. A method of generating purified oligonucleotides comprising:
-
a) providing;
i) a substrate comprising an array of oligonucleotides attached to said substrate via anchor moieties attached to non-cleavable linkers attached to said substrate, wherein said anchor moieties comprise the structure —
C(X)—
C(Y), wherein X comprises —
OPO2O—
, Y is a nucleophile and wherein said structure is part of a ring moiety;
ii) a deprotecting solution; and
iii) and a wash solution;
b) deprotecting said oligonucleotides with said deprotecting solution;
c) washing said oligonucleotides with said wash solution; and
d) cleaving said oligonucleotides at said anchor group to provide purified oligonucleotides. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19)
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Specification