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US 12,168,211 B2Granted
Reusable initiators for synthesizing nucleic acids
AssigneeMolecular Assemblies, Inc.
InventorsJ. William Efcavitch, Matthew T. Holden
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Abstract
The invention provides improved methods for synthesizing polynucleotides, such as DNA and RNA, using renewable initiators coupled to a solid support. Using the methods of the invention, specific sequences of polynucleotides can be synthesized de novo, base by base, in an aqueous environment, without the use of a nucleic acid template.
Claims13
A method for synthesizing oligonucleotides on a reusable solid support, the method comprising:
- exposing a nucleic acid initiator attached to a solid support to nucleotide analogs in the presence of a polymerase to create a first oligonucleotide, wherein the nucleic acid initiator comprises a sequence-specific cleavage element;
- contacting the nucleic acid initiator with a releasing agent to cleave the sequence-specific cleavage element and release the first oligonucleotide from the nucleic acid initiator;
- dephosphorylating a 3′-end of the solid support-attached nucleic acid initiator;
- and
- enzymatically renewing the sequence-specific cleavage element.
2 more elements
Dependencies are inferred from the claim text.