公开(公告)号：US10689680B2

公开(公告)日：2020-06-23

申请号：US15953784

申请日：2018-04-16

Applicant: Agilent Technologies, Inc.

Inventor： Jeffrey Robert Sampson ,  Derek Lee Lindstrom

IPC: C12P19/34 ,  C12N15/10 ,  C12N15/64 ,  C12N15/66

Abstract: Methods for making a synthetic nucleic acid which comprise: (a) identifying a conflicting nucleotide sequence in a target sequence; (b) inserting a masking sequence into the conflicting sequence to produce a disrupted target sequence, wherein: (i) the masking sequence comprises recognition sites for one or more Type IIS restriction endonucleases; and (ii) digestion of said disrupted target sequence by said one or more Type IIS restriction endonucleases followed by re-ligation reconstitutes the target sequence; (c) synthesizing a polynucleotide comprising the disrupted target sequence using polymerase chain assembly; and (d) removing the masking sequence from said polynucleotide by digesting said polynucleotide with said one or more Type IIS restriction endonucleases followed by re-ligation of the digestion product, thereby producing a polynucleotide comprising said target sequence.

公开(公告)号：US20180237818A1

公开(公告)日：2018-08-23

申请号：US15953784

申请日：2018-04-16

Applicant: Agilent Technologies, Inc.

Inventor： Jeffrey Robert Sampson ,  Derek Lee Lindstrom

IPC: C12P19/34 ,  C12N15/66 ,  C12N15/10 ,  C12N15/64

CPC classification number: C12P19/34 ,  C12N15/10 ,  C12N15/64 ,  C12N15/66

Abstract: Methods for making a synthetic nucleic acid which comprise: (a) identifying a conflicting nucleotide sequence in a target sequence; (b) inserting a masking sequence into the conflicting sequence to produce a disrupted target sequence, wherein: (i) the masking sequence comprises recognition sites for one or more Type IIS restriction endonucleases; and (ii) digestion of said disrupted target sequence by said one or more Type IIS restriction endonucleases followed by re-ligation reconstitutes the target sequence; (c) synthesizing a polynucleotide comprising the disrupted target sequence using polymerase chain assembly; and (d) removing the masking sequence from said polynucleotide by digesting said polynucleotide with said one or more Type IIS restriction endonucleases followed by re-ligation of the digestion product, thereby producing a polynucleotide comprising said target sequence.

公开(公告)号：US09976162B2

公开(公告)日：2018-05-22

申请号：US14649111

申请日：2013-11-05

Applicant: Agilent Technologies, Inc.

Inventor： Jeffrey Robert Sampson ,  Derek Lee Lindstrom

IPC: C12P19/34 ,  C12N15/64 ,  C12N15/10 ,  C12N15/66

CPC classification number: C12P19/34 ,  C12N15/10 ,  C12N15/64 ,  C12N15/66

Abstract: Provided herein is a method comprising: producing a first complex by annealing: a first nucleic acid comprising, in order, a first unique sequence, a first central sequence and a second unique sequence; and a second nucleic acid comprising, in order, said second unique sequence, a second central sequence and a third unique sequence; wherein the first, second and third unique sequences do not hybridize with each other; subjecting said first complex to multiple rounds of primer extension to extend the first and second nucleic acids using each other as a template, thereby producing a first product molecule that contains, in order, the first unique sequence, the first central sequence, the second unique sequence, the second central sequence and the third unique sequence; and circularizing said first product molecule by intramolecularly ligating the ends of said product molecule together. Kits and compositions relating to the method are also provided.

公开(公告)号：US10308979B2

公开(公告)日：2019-06-04

申请号：US13830408

申请日：2013-03-14

Applicant: Agilent Technologies, Inc.

Inventor： Brian Jon Peter ,  Robert A. Ach ,  Zoltan Timar ,  Joel Myerson ,  Jeffrey Robert Sampson ,  Holly Hogrefe

IPC: C12Q1/68 ,  C12Q1/6813 ,  C12Q1/6806

Abstract: This disclosure provides a method comprising: a) clamping the top and bottom strands of a double stranded DNA molecule to produce a duplex in which the top and bottom strands are linked; b) denaturing the duplex to produce a denatured product; and c) renaturing the denatured product in the presence of a labeled oligonucleotide that is complementary to a sequence of nucleotides in the double stranded DNA molecule, thereby producing a D-loop-containing product. Kits for performing the method and products made by the method are also provided.

公开(公告)号：US20150344927A1

公开(公告)日：2015-12-03

申请号：US14649111

申请日：2013-11-05

Applicant: AGILENT TECHNOLOGIES, INC.

Inventor： Jeffrey Robert Sampson ,  Derek Lee Lindstrom

IPC: C12P19/34 ,  C12N15/64

CPC classification number: C12P19/34 ,  C12N15/10 ,  C12N15/64 ,  C12N15/66

Abstract: Provided herein is a method comprising: producing a first complex by annealing: a first nucleic acid comprising, in order, a first unique sequence, a first central sequence and a second unique sequence; and a second nucleic acid comprising, in order, said second unique sequence, a second central sequence and a third unique sequence; wherein the first, second and third unique sequences do not hybridize with each other; subjecting said first complex to multiple rounds of primer extension to extend the first and second nucleic acids using each other as a template, thereby producing a first product molecule that contains, in order, the first unique sequence, the first central sequence, the second unique sequence, the second central sequence and the third unique sequence; and circularizing said first product molecule by intramolecularly ligating the ends of said product molecule together. Kits and compositions relating to the method are also provided.

Abstract translation: 本文提供的方法包括：通过退火产生第一配合物：第一核酸，依次包含第一唯一序列，第一中心序列和第二唯一序列; 和第二核酸，依次包含所述第二唯一序列，第二中心序列和第三唯一序列; 其中所述第一，第二和第三独特序列彼此不杂交; 使所述第一复合物经受多轮引物延伸以使第一和第二核酸彼此作为模板，从而产生第一产物分子，依次包含第一独特序列，第一中心序列，第二独特序列 序列，第二中心序列和第三个唯一序列; 并通过将所述产物分子的末端分子内连接在一起使所述第一产物分子环化。 还提供了与该方法相关的试剂盒和组合物。