Methods for in situ generation of nucleic acid molecules
First Claim
1. A method for synthesizing a nucleic acid molecule on a substrate, comprising:
- contacting a substrate comprising an attached blocked nucleoside monomer or polymer with a deblocking fluid to remove the blocking group, thereby generating an unblocked attached nucleoside monomer or polymer;
displacing the deblocking fluid from the substrate surface comprising the attached unblocked nucleoside monomer or polymer with a purging fluid;
reacting the attached unblocked nucleoside monomer or polymer with another blocked nucleoside monomer.
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Abstract
Methods of producing nucleic acid molecules using an in situ nucleic acid synthesis protocol are provided. The method can comprise contacting a substrate comprising an attached blocked nucleoside monomer or polymer with a deblocking fluid to remove the blocking group, thereby generating an unblocked attached nucleoside monomer or polymer; displacing the deblocking fluid from the substrate surface comprising the attached unblocked nucleoside monomer or polymer with a purging fluid; and reacting the attached unblocked nucleoside monomer or polymer with another blocked nucleoside monomer. Nucleic acid molecules produced by the methods are also provided and can be attached to or released from the substrate (e.g., provided in solution or in a lyophilized form).
67 Citations
46 Claims
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1. A method for synthesizing a nucleic acid molecule on a substrate, comprising:
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contacting a substrate comprising an attached blocked nucleoside monomer or polymer with a deblocking fluid to remove the blocking group, thereby generating an unblocked attached nucleoside monomer or polymer;
displacing the deblocking fluid from the substrate surface comprising the attached unblocked nucleoside monomer or polymer with a purging fluid;
reacting the attached unblocked nucleoside monomer or polymer with another blocked nucleoside monomer. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 38, 39, 40, 41, 42, 43, 44, 45, 46)
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33. A method of producing a substrate of at least two oligonucleotides bonded to different locations on a surface of the substrate, comprising:
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contacting blocked nucleoside monomers to at least a first location and a second different location of a substrate surface displaying functional groups under conditions sufficient for the blocked nucleoside monomers to bond to the surface in first and second locations to produce a substrate surface displaying bound blocked monomers;
contacting the surface displaying bound blocked monomers with a deblocking fluid to remove the blocking group, thereby generating unblocked nucleoside monomers at the first and second locations;
displacing the deblocking fluid from the surface comprising the bound unblocked monomers at the first and second locations with a purging fluid;
reacting the attached unblocked nucleoside monomers at the first and second locations with another blocked nucleoside monomer. - View Dependent Claims (34, 35, 36, 37)
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Specification