Covalent attachment of nucleic acid molecules onto solid-phases via disulfide bonds
First Claim
1. A method for attaching a modified nucleic acid molecule to a coated solid-phase via a disulfide bond, said modified nucleic acid molecule being modified such that a 3'"'"' carbon atom or a 5'"'"' carbon atom of a sugar moiety of a nucleotide of said nucleic acid molecule possesses a sulfhydryl group or a disulfide group, said method comprising coupling said modified nucleic acid molecule to said coated solid-phase, wherein where said modified nucleic acid molecule is a modified nucleic acid molecule having a sulfhydryl group, said coated solid-phase is coated with a disulfidesilane, and where said modified nucleic acid molecule is a modified nucleic acid molecule having a disulfide group said coated solid-phase is coated with a mercaptosilane and wherein said disulfide bond is formed between a sulfur atom of either said sulfhydryl group or said disulfide group and a sulfur atom of either said disulfidesilane or said mercaptosilane.
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Accused Products
Abstract
Methods for the covalent, specific and reversible immobilization of nucleic acid molecules onto solid-phases by means of a reversible disulfide bond for nucleic acid molecule array preparation are described. These methods can be used to prepare reusable nucleic acid molecule arrays with hilgh specificity and high efficiency.
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Citations
31 Claims
- 1. A method for attaching a modified nucleic acid molecule to a coated solid-phase via a disulfide bond, said modified nucleic acid molecule being modified such that a 3'"'"' carbon atom or a 5'"'"' carbon atom of a sugar moiety of a nucleotide of said nucleic acid molecule possesses a sulfhydryl group or a disulfide group, said method comprising coupling said modified nucleic acid molecule to said coated solid-phase, wherein where said modified nucleic acid molecule is a modified nucleic acid molecule having a sulfhydryl group, said coated solid-phase is coated with a disulfidesilane, and where said modified nucleic acid molecule is a modified nucleic acid molecule having a disulfide group said coated solid-phase is coated with a mercaptosilane and wherein said disulfide bond is formed between a sulfur atom of either said sulfhydryl group or said disulfide group and a sulfur atom of either said disulfidesilane or said mercaptosilane.
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2. A method for attaching a nucleic acid molecule to a solid-phase via a disulfide bond, said nucleic acid molecule being modified such that a 3'"'"' carbon atom or a 5'"'"' carbon atom of a sugar moiety of a nucleotide of said nucleic acid molecule possesses a sulthydryl group or a disulfide group, said method comprising the steps of:
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(A) coating the surface of the solid-phase with a mercaptosilane or a disulfidesilane, wherein said mercaptosilane comprises a sulfliydryl group and said disulfidesilane comprise a disulfide group; and (B) (i) coupling a sulfhydryl modified nucleic acid molecule to the disulfide group of said disulfidesilane by means of a covalent disulfide bond;
or(ii) coupling a disulfide modified nucleic acid molecule to the sulfhydryl groups of said mercaptosilane by means of a covalent disulfide bond; wherein said disulfide bond is formed between a sulfur atom of either said sulfhydryl group or said disulfide group and a sulfur atom of either said disulfidesilane or said mercaptosilane. - View Dependent Claims (17, 18, 19, 23)
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31. A modified nucleic acid molecule coupled via a disulfide bond to a coated solid-phase, said modified nucleic acid molecule being modified such that a 3'"'"' carbon atom or a 5'"'"' carbon atom of a sugar moiety of a nucleotide of said nucleic acid molecule possesses a sulfhydryl group or a disulfide group, wherein where said modified nucleic acid molecule is a modified nucleic acid molecule having a sulfhydryl group, said coated solid-phase is coated with a disulfidesilane, and where said modified nucleic acid molecule is a modified nucleic acid molecule having a disulfide group said coated solid-phase is coated with a mercaptosilane, and wherein said disulfide bond is formed between a sulfur atom of either said sulfhydryl group or said disulfide group and a sulfur atom of either said disulfidesilane or said mercaptosilane.
Specification