公开(公告)号：US10294514B2

公开(公告)日：2019-05-21

申请号：US15581828

申请日：2017-04-28

Applicant: Omniome, Inc.

Inventor： Pinar Iyidogan ,  Kandaswamy Vijayan

IPC: C12Q1/68 ,  C12Q1/6809 ,  C12Q1/6816 ,  C12Q1/6848 ,  C12Q1/6869 ,  C12Q1/6874

Abstract: Provided are methods and systems for detecting formation of nucleotide-specific ternary complexes comprising a DNA polymerase, a nucleic acid, and a nucleotide complementary to the templated base of the primed template nucleic acid. The methods and systems facilitate determination of the next correct nucleotide without requiring chemical incorporation of the nucleotide into the primer. These results can even be achieved in procedures employing unlabeled, native nucleotides.

公开(公告)号：US11203778B2

公开(公告)日：2021-12-21

申请号：US16823610

申请日：2020-03-19

Applicant: Omniome, Inc.

Inventor： Pinar Iyidogan ,  Kandaswamy Vijayan

IPC: C12Q1/6809 ,  C12Q1/6874 ,  C12Q1/6816 ,  C12Q1/6848 ,  C12Q1/6869

Abstract: Provided are methods and systems for detecting formation of nucleotide-specific ternary complexes comprising a DNA polymerase, a nucleic acid, and a nucleotide complementary to the templated base of the primed template nucleic acid. The methods and systems facilitate determination of the next correct nucleotide without requiring chemical incorporation of the nucleotide into the primer. These results can even be achieved in procedures employing unlabeled, native nucleotides.

公开(公告)号：US10731141B2

公开(公告)日：2020-08-04

申请号：US16567598

申请日：2019-09-11

Applicant: OMNIOME, INC.

Inventor： Pinar Iyidogan

IPC: C12N9/12 ,  C12P19/34 ,  C12Q1/6874

Abstract: Provided are nucleic acids encoding engineered polymerases comprising at least one modification in a motif A and/or at least one modification in a motif B of the polymerase and engineered polymerases encoded by the nucleic acids. Also provided are engineered DNA polymerases comprising a variant of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3, the variant being at least 80% identical to SEQ ID NO:1, SEQ ID NO:2 or SEQ ID NO:3 and comprising an amino acid substitution at one or more positions selected from the group consisting of L408, Y409, P410, R484, A/L485, and I486. Methods, vectors, kits, and compositions comprising the nucleic acids and compositions, methods and kits comprising the engineered polymerases are also provided.

公开(公告)号：US10597643B2

公开(公告)日：2020-03-24

申请号：US15866353

申请日：2018-01-09

Applicant: OMNIOME, INC.

Inventor： Pinar Iyidogan ,  Mark C. Wallen ,  Ying L. Liu ,  Kandaswamy Vijayan

IPC: C12N9/12 ,  C12Q1/6869

Abstract: Provided are engineered DNA polymerases exhibiting modified functionality, and polynucleotides encoding same. Modified features include: (1) reduced catalytic activity in the presence of magnesium ions and/or (2) reduced affinity for primed template nucleic acid molecules in the absence of cognate nucleotide, and an ability to discriminate between cognate and non-cognate nucleotides under low salt conditions. Sequencing By Binding™ procedures employing the engineered polymerases have certain advantages. The engineered polymerases can have other uses as well.

公开(公告)号：US10400272B1

公开(公告)日：2019-09-03

申请号：US16355361

申请日：2019-03-15

Applicant: Omniome, Inc.

Inventor： Morassa Mohseni Middleton ,  Mark C. Wallen ,  Pinar Iyidogan ,  Michael James Schmidt ,  Brittany A. Rohrman ,  Ying Lin Liu ,  Fabian Block ,  Arnold Oliphant

IPC: C12Q1/68 ,  C12Q1/6832 ,  C12Q1/6816

Abstract: Methods, compositions, kits and apparatuses that include a fluid, the fluid containing a ternary complex and Li+, wherein the ternary complex includes a primed template nucleic acid, a polymerase, and a nucleotide cognate for the next correct base for the primed template nucleic acid molecule. As an alternative or addition to Li+, the fluid can contain betaine or a metal ion that inhibits polymerase catalysis such as Ca2+. In addition to Li+, the fluid can contain polyethylenimine (PEI) with or without betaine.

公开(公告)号：US20180044715A1

公开(公告)日：2018-02-15

申请号：US15654406

申请日：2017-07-19

Applicant: Omniome, Inc.

Inventor： Pinar Iyidogan ,  Kandaswamy Vijayan

IPC: C12Q1/68 ,  G01N27/447

CPC classification number: C12Q1/686 ,  C12Q1/00 ,  C12Q1/6806 ,  C12Q1/6813 ,  C12Q1/6853 ,  C12Q1/6869 ,  G01N27/447 ,  G01N2001/4038 ,  C12Q2525/186 ,  C12Q2535/113 ,  C12Q2565/133 ,  C12Q2565/607 ,  C12Q2565/628

Abstract: Provided are methods and systems for reducing the time needed for sequencing nucleic acids. The approach relies on detecting formation of nucleotide-specific ternary complexes comprising a polymerase (e.g., a DNA polymerizing enzyme), a primed template nucleic acid molecule, and a nucleotide complementary to the templated base of the primed template nucleic acid. The methods and systems facilitate determination of the next correct nucleotide, as well as the subsequent next correct nucleotide from a cycle of examining four different nucleotides without requiring chemical incorporation of any nucleotide into the primer.

公开(公告)号：US20170314072A1

公开(公告)日：2017-11-02

申请号：US15581822

申请日：2017-04-28

Applicant: Omniome, Inc.

Inventor： Kandaswamy Vijayan ,  Pinar Iyidogan

IPC: C12Q1/68 ,  C12N9/12

Abstract: Provided are sequencing-by-binding methods of detecting cognate nucleotides using a crippled DNA polymerizing enzyme that possesses the ability to bind the next correct nucleotide downstream of a primer in a template-dependent fashion, but does not possess the activity needed to promote phosphodiester bond formation. Use of the crippled DNA polymerase permits interrogation of one nucleotide at a time, without incorporation of any nucleotide. Labeled nucleotides, such as fluorescently labeled nucleotides, can be used in conjunction with the crippled DNA polymerase to establish cognate nucleotide identity in a rapid manner.

公开(公告)号：US11242557B2

公开(公告)日：2022-02-08

申请号：US16516808

申请日：2019-07-19

Applicant: Omniome, Inc.

Inventor： Morassa Mohseni Middleton ,  Mark C. Wallen ,  Pinar Iyidogan ,  Michael James Schmidt ,  Brittany A. Rohrman ,  Ying Lin Liu ,  Fabian Block ,  Arnold Oliphant

IPC: C12Q1/6832 ,  C12Q1/6816

Abstract: Methods, compositions, kits and apparatuses that include a fluid, the fluid containing a ternary complex and Li+, wherein the ternary complex includes a primed template nucleic acid, a polymerase, and a nucleotide cognate for the next correct base for the primed template nucleic acid molecule. As an alternative or addition to Li+, the fluid can contain betaine or a metal ion that inhibits polymerase catalysis such as Ca2+. In addition to Li+, the fluid can contain polyethylenimine (PEI) with or without betaine.

公开(公告)号：US10584379B2

公开(公告)日：2020-03-10

申请号：US15581822

申请日：2017-04-28

Applicant: Omniome, Inc.

Inventor： Kandaswamy Vijayan ,  Pinar Iyidogan

IPC: C12Q1/68 ,  C12Q1/6869 ,  C12Q1/6874 ,  C12N9/12

Abstract: Provided are sequencing-by-binding methods of detecting cognate nucleotides using a crippled DNA polymerizing enzyme that possesses the ability to bind the next correct nucleotide downstream of a primer in a template-dependent fashion, but does not possess the activity needed to promote phosphodiester bond formation. Use of the crippled DNA polymerase permits interrogation of one nucleotide at a time, without incorporation of any nucleotide. Labeled nucleotides, such as fluorescently labeled nucleotides, can be used in conjunction with the crippled DNA polymerase to establish cognate nucleotide identity in a rapid manner.

公开(公告)号：US20190226012A1

公开(公告)日：2019-07-25

申请号：US16372812

申请日：2019-04-02

Applicant: Omniome, Inc.

Inventor： Pinar Iyidogan ,  Kandaswamy Vijayan

IPC: C12Q1/6809 ,  C12Q1/6874 ,  C12Q1/6816 ,  C12Q1/6848 ,  C12Q1/6869

Abstract: Provided are methods and systems for detecting formation of nucleotide-specific ternary complexes comprising a DNA polymerase, a nucleic acid, and a nucleotide complementary to the templated base of the primed template nucleic acid. The methods and systems facilitate determination of the next correct nucleotide without requiring chemical incorporation of the nucleotide into the primer. These results can even be achieved in procedures employing unlabeled, native nucleotides.