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US 2021/0261999 A1Published application
Solid-phase synthesis of polynucleotides using a template array
AssigneeTemplanucleics, Inc.
InventorsRichard FUISZ, Alan TOMUSIAK
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Abstract
Provided herein are methods and systems for synthesizing polynucleotides using a solid support and a template array. Also provided herein are methods of selecting templates for the array.
Claims20
A method of extending a primer to generate a polynucleotide, comprising:
- (a) providing a primer, wherein the primer has a 5′ fixed end and a 3′ growing end, wherein the fixed end is bound to a solid substrate;
- (b) providing an array comprising a plurality of template wells, wherein at least a portion of the wells each comprise two or more unique templates in solution;
- (c) contacting the solid substrate with the solution of a well comprising two or more unique templates,
- wherein one of the two or more unique templates is a complementary template comprising a complementary region, wherein the sequence of the complementary region is complementary to the growing end;
- and
- at least one of the two or more unique templates is a non-complementary template, wherein the sequence of the non-complementary template is not complementary to the growing end;
- (d) hybridizing the growing end and the complementary region to provide an overhang of the complementary template comprising one or more nucleotides;
- (e) extending the growing end of the primer by the addition of one or more nucleotides, wherein the one or more added nucleotides are complementary to the one or more nucleotides of the overhang region;
- (f) dehybridizing the growing end and the complementary region;
- and
- (g) repeating steps (c) through (f) to generate the polynucleotide.
8 more elements
A system for extending a growing primer to generate a polynucleotide, comprising:
- a growing primer comprising a 5′ fixed end and a 3′ growing end, wherein the fixed end is bound to a solid substrate;
- a template array comprising a plurality of template wells, wherein at least a portion of the template wells each comprise two or more unique templates in solution,
- wherein in one of the template wells, one of the two or more unique templates is a complementary template comprising a complementary region, wherein the sequence of the complementary region is complementary to the growing end;
- and
- at least one of the two or more unique templates is a non-complementary template, wherein the sequence of the non-complementary template is not complementary to the growing end;
- and
- a positioning apparatus configured to contact the solid substrate with the solution of a well.
4 more elements
Dependencies are inferred from the claim text.