Synthesis of long nucleic acid sequences

Patent metadata

Field	Value
Publication number	US-11034989-B2
Application number	US-201715591783-A
Country	US
Kind code	B2
Filing date	May 10, 2017
Priority date	Jan 16, 2012
Publication date	Jun 15, 2021
Grant date	Jun 15, 2021

Abstract

The invention provides methods for the synthesis of long oligonucleotides, genes and gene fragments. The methods include the manufacture of genes or gene fragments that can be then inserted into a variety of vectors.

First claim

What is claimed is:

1. A gene fragment library, said library comprising: a plurality of non-vector synthetic nucleic acid gene fragments wherein two or more of the gene fragments are comprised of a constant gene block a of at least 100 bases and a variable gene block of at least 50 bases, wherein the constant gene block sequence is identical for each of the gene fragments and the variable gene block sequence varies, and wherein the plurality of non-vector synthetic nucleic acid gene fragments are chemically synthesized.

2. The gene fragment library of claim 1 wherein the gene fragments further comprise constant gene block b .

3. The gene fragment library of claim 2 wherein the variable gene block is flanked by constant gene block a and constant gene block b .

4. The gene fragment library of claim 2 wherein the gene fragments further comprise constant gene block n wherein n represents a plurality of sets of constant gene blocks.

5. The gene fragment library of claim 1 wherein the gene fragments further comprise variable gene block n wherein n represents a plurality of sets of variable gene blocks.

6. The gene fragment library of claim 1 wherein the gene fragments are at least 400 bases.

7. The gene fragment library of claim 1 wherein the gene fragments are at least 1000 bases.

8. The gene fragment library of claim 3 wherein the constant gene blocks located on terminal ends contain a binding site for a forward primer on a first gene block located at a first terminal end and a reverse amplification primer on a second gene block located at a second terminal end.

9. The gene fragment library of claim 1 wherein the constant gene blocks and the variable gene blocks contain overlap regions for assembly into a gene fragment.

10. The gene fragment library of claim 9 wherein the overlap regions contain a same sequence of complementarity that allow for assembly with universal primers.

Assignees

• Integrated Dna Tech Inc

Inventors

• Allen Shawn
• Gunstream Stephen
• Rose Scott

Classifications

• C12P19/34 Primary

  Polynucleotides, e.g. nucleic acids, oligoribonucleotides · CPC title

• B01J2219/00722

  Nucleotides · CPC title

• B01J2219/00596

  Solid-phase processes · CPC title

• C12N15/1068

  Template (nucleic acid) mediated chemical library synthesis, e.g. chemical and enzymatical DNA-templated organic molecule synthesis, libraries prepared by non ribosomal polypeptide synthesis [NRPS], DNA/RNA-polymerase mediated polypeptide synthesis · CPC title

• C12Q1/6844

  Nucleic acid amplification reactions · CPC title