US 12,168,211 B2
Reusable initiators for synthesizing nucleic acids
J. William Efcavitch, San Carlos, CA (US); and Matthew T. Holden, San Diego, CA (US)
Assigned to Molecular Assemblies, Inc., San Diego, CA (US)
Filed by Molecular Assemblies, Inc., San Diego, CA (US)
Filed on May 16, 2022, as Appl. No. 17/745,128.
Application 17/745,128 is a continuation of application No. 16/261,229, filed on Jan. 29, 2019, granted, now 11,331,643.
Application 16/261,229 is a continuation in part of application No. 14/829,269, filed on Aug. 18, 2015, granted, now 10,683,536, issued on Jun. 16, 2020.
Application 14/829,269 is a continuation in part of application No. 14/459,014, filed on Aug. 13, 2014, granted, now 9,279,149, issued on Mar. 8, 2016.
Application 14/459,014 is a continuation in part of application No. 14/056,687, filed on Oct. 17, 2013, granted, now 8,808,989, issued on Aug. 19, 2014.
Claims priority of provisional application 62/069,067, filed on Oct. 27, 2014.
Claims priority of provisional application 62/038,604, filed on Aug. 18, 2014.
Claims priority of provisional application 61/891,162, filed on Oct. 15, 2013.
Claims priority of provisional application 61/807,327, filed on Apr. 2, 2013.
Prior Publication US 2023/0103381 A1, Apr. 6, 2023
Int. Cl. C12Q 1/68 (2018.01); B01J 19/00 (2006.01); C12N 9/12 (2006.01); C12P 19/34 (2006.01); C12Q 1/6806 (2018.01); C12Q 1/6811 (2018.01); C12Q 1/6837 (2018.01)
CPC B01J 19/0046 (2013.01) [C12N 9/1264 (2013.01); C12P 19/34 (2013.01); C12Q 1/6806 (2013.01); C12Q 1/6811 (2013.01); C12Q 1/6837 (2013.01); B01J 2219/00596 (2013.01); B01J 2219/00722 (2013.01); C12Q 2525/101 (2013.01); C12Q 2525/117 (2013.01); C12Q 2525/186 (2013.01); C12Y 207/07019 (2013.01); C12Y 207/07031 (2013.01)] 13 Claims
 
1. 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.