公开(公告)号：US20070020669A1

公开(公告)日：2007-01-25

申请号：US11483825

申请日：2006-07-10

申请人： Olof Ericsson

发明人： Olof Ericsson

IPC分类号： C12Q1/68 ,  G06F19/00

CPC分类号： C12Q1/6804 ,  C12N15/1075 ,  G01N33/53 ,  G01N33/58 ,  G01N2458/10

摘要： Methods of detecting affinity interactions between at least two molecules of interest are provided. The method comprises: a. forming a plurality of interactors by coupling each molecule of interest with at least one nucleic acid moiety comprising an identification sequence element and at an association element; b. promoting an association between at least two nucleic acid moieties from different interactors to form a plurality of unique associated oligonucleotides, wherein each nucleic acid moiety may form more than one unique associated oligonucleotide, and wherein each unique associated oligonucleotide comprises at least two identification sequence elements derived from the at least two nucleic acid moieties; c. selecting the plurality of unique associated oligonucleotides; and d. subjecting the selected associated oligonucleotides to an analysis that permits detection of the at least two identification sequence elements. Similar methods directed to detecting functional interactions, libraries of interactors employable in the present methods, and kits comprising those libraries are also provided.

摘要翻译： 提供了检测至少两个感兴趣的分子之间的亲和力相互作用的方法。 该方法包括：a。 通过将每个感兴趣的分子与至少一个包含识别序列元件的核酸部分和缔合元件相耦合来形成多个相互作用体; b。 促进来自不同相互作用者的至少两个核酸部分之间的缔合以形成多个独特的相关寡核苷酸，其中每个核酸部分可以形成多于一个唯一相关的寡核苷酸，并且其中每个唯一相关寡核苷酸包含至少两个识别序列元件 从所述至少两个核酸部分; C。 选择所述多个独特的相关寡核苷酸; 和d。 对所选择的相关寡核苷酸进行允许检测所述至少两个识别序列元件的分析。 还提供了用于检测功能相互作用的类似方法，在本方法中可使用的相互作用者的文库以及包括这些文库的试剂盒。

公开(公告)号：US20120289426A1

公开(公告)日：2012-11-15

申请号：US13512924

申请日：2010-12-03

申请人： Fredrik Roos ,  Henrik Johansson ,  Magnus Isaksson ,  Mats Nilsson ,  Olle (Olof) Ericsson ,  Simon Fredriksson

发明人： Fredrik Roos ,  Henrik Johansson ,  Magnus Isaksson ,  Mats Nilsson ,  Olle (Olof) Ericsson ,  Simon Fredriksson

IPC分类号： C40B30/04 ,  C40B50/06 ,  C12Q1/68

CPC分类号： C12Q1/6837 ,  C12Q2565/519 ,  C12Q2531/125 ,  C12Q2563/131 ,  C12Q2525/161 ,  C12Q2521/501

摘要： A method for amplifying a target nucleic acid is disclosed, which includes: (a) fragmenting a nucleic acid sample to create a target fragment comprising a target nucleic acid and two probe-complementary portions; (b) contacting said fragmented nucleic acid sample with a probe comprising two target fragment-complementary portions complementary to the probe-complementary portions of the target fragment; (c) rendering the fragmented nucleic acid sample single-stranded; (d) allowing the probe-complementary portions to hybridise with the target-fragment complementary portions; (e) if the probe in step (b) is not immobilised, immobilising the probe-target fragment hybrid on a solid phase via immobilisation moiety; (f) separating non-immobilised nucleic acid fragments from the solid phase; (g) contacting the solid phase with a ligase to ligate ligatable 5′ and 3′ ends of the target fragment whereby the target fragment is circularized; and (h) amplifying said circularized target fragment.

摘要翻译： 公开了扩增靶核酸的方法，其包括：（a）将核酸样品片段化以产生包含靶核酸和两个探针互补部分的靶片段; （b）使所述片段化的核酸样品与包含与靶片段的探针互补部分互补的两个靶片段互补部分的探针接触; （c）使片段化的核酸样品单链; （d）允许探针互补部分与靶片段互补部分杂交; （e）如果步骤（b）中的探针未固定，则通过固定部分将探针 - 靶片段杂交体固定在固相上; （f）从固相中分离未固定的核酸片段; （g）使固相与连接酶接触以连接靶片段的可连接的5'和3'末端，由此靶片段被环化; 和（h）扩增所述环化的靶片段。

公开(公告)号：US20120220479A1

公开(公告)日：2012-08-30

申请号：US13384990

申请日：2010-07-23

申请人： Olof Ericsson ,  Magnus Isaksson ,  Henrik Johansson ,  Ulf Landegren

发明人： Olof Ericsson ,  Magnus Isaksson ,  Henrik Johansson ,  Ulf Landegren

IPC分类号： C12Q1/68 ,  C40B30/04

CPC分类号： C12Q1/6855

摘要： The present invention provides a method for detecting or enriching for a target deoxyribonucleic acid (DNA) present in a nucleic acid sample, said method comprising: (a) fragmenting a nucleic acid sample to generate nucleic acid fragments including a target fragment containing said target DNA; (b) rendering said fragments, including said target fragment, at least partially single-stranded, wherein the single-stranded portion includes an end portion and wherein the length of said single-stranded portion is sufficient to allow hybridisation of at least part of the single-stranded portion of said target fragment to the probe of step (c); (c) contacting the at least partially single-stranded fragments of step (b) with oligonucleotides A and B of a single target-specific nucleic acid probe, wherein: (i) oligonucleotide A is a single-stranded oligonucleotide comprising at one end a first target-specific part comprising at least 10 nucleotides complementary in sequence to at least part of said single-stranded portion of said target fragment, and comprising at the other end a second non-target-specific part which comprises a nucleotide sequence complementary to at least a portion, including one end, of oligonucleotide B of the probe, and (ii) oligonucleotide B is a single-stranded oligonucleotide which may contain or carry at least one element for detection and/or enrichment of said target fragment, and of which at least a portion, including one end, is complementary in sequence to the second non-target-specific part of oligonucleotide A, such that said target fragment becomes annealed to said probe through hybridisation to the first target-specific part of oligonucleotide A resulting in only one target-specific probe-binding event per target fragment; (d) ligating oligonucleotide B of said probe to the part of the single-stranded portion of said target fragment which is hybridised to oligonucleotide A of said probe to produce a probe-target fragment hybrid; and (e) detecting or enriching for said probe-target fragment hybrid. Kits for use in the method of the invention are also provided.

公开(公告)号：US07883848B2

公开(公告)日：2011-02-08

申请号：US11483825

申请日：2006-07-10

申请人： Olof Ericsson

发明人： Olof Ericsson

IPC分类号： C12Q1/68 ,  C12P19/34 ,  C07H21/02 ,  G01N33/566

CPC分类号： C12Q1/6804 ,  C12N15/1075 ,  G01N33/53 ,  G01N33/58 ,  G01N2458/10

摘要： Methods of detecting affinity interactions between at least two molecules of interest are provided. The method comprises: a. forming a plurality of interactors by coupling each molecule of interest with at least one nucleic acid moiety comprising an identification sequence element and at an association element; b. promoting an association between at least two nucleic acid moieties from different interactors to form a plurality of unique associated oligonucleotides, wherein each nucleic acid moiety may form more than one unique associated oligonucleotide, and wherein each unique associated oligonucleotide comprises at least two identification sequence elements derived from the at least two nucleic acid moieties; c. selecting the plurality of unique associated oligonucleotides; and d. subjecting the selected associated oligonucleotides to an analysis that permits detection of the at least two identification sequence elements. Similar methods directed to detecting functional interactions, libraries of interactors employable in the present methods, and kits comprising those libraries are also provided.

摘要翻译： 提供了检测至少两个感兴趣的分子之间的亲和力相互作用的方法。 该方法包括：a。 通过将每个感兴趣的分子与至少一个包含识别序列元件的核酸部分和缔合元件相耦合来形成多个相互作用体; b。 促进来自不同相互作用者的至少两个核酸部分之间的缔合以形成多个独特的相关寡核苷酸，其中每个核酸部分可以形成多于一个唯一相关的寡核苷酸，并且其中每个唯一相关寡核苷酸包含至少两个识别序列元件 从所述至少两个核酸部分; C。 选择所述多个独特的相关寡核苷酸; 和d。 对所选择的相关寡核苷酸进行允许检测所述至少两个识别序列元件的分析。 还提供了用于检测功能相互作用的类似方法，在本方法中可使用的相互作用者的文库以及包括这些文库的试剂盒。

公开(公告)号：US09657339B2

公开(公告)日：2017-05-23

申请号：US13512924

申请日：2010-12-03

申请人： Fredrik Roos ,  Henrik Johansson ,  Magnus Isaksson ,  Mats Nilsson ,  Olle (Olof) Ericsson ,  Simon Fredriksson

发明人： Fredrik Roos ,  Henrik Johansson ,  Magnus Isaksson ,  Mats Nilsson ,  Olle (Olof) Ericsson ,  Simon Fredriksson

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6837 ,  C12Q2565/519 ,  C12Q2531/125 ,  C12Q2563/131 ,  C12Q2525/161 ,  C12Q2521/501

摘要： A method for amplifying a target nucleic acid is disclosed, which includes: (a) fragmenting a nucleic acid sample to create a target fragment comprising a target nucleic acid and two probe-complementary portions; (b) contacting said fragmented nucleic acid sample with a probe comprising two target fragment-complementary portions complementary to the probe-complementary portions of the target fragment; (c) rendering the fragmented nucleic acid sample single-stranded; (d) allowing the probe-complementary portions to hybridise with the target-fragment complementary portions; (e) if the probe in step (b) is not immobilised, immobilising the probe-target fragment hybrid on a solid phase via immobilisation moiety; (f) separating non-immobilised nucleic acid fragments from the solid phase; (g) contacting the solid phase with a ligase to ligate ligatable 5′ and 3′ ends of the target fragment whereby the target fragment is circularized; and (h) amplifying said circularized target fragment.

公开(公告)号：US08664164B2

公开(公告)日：2014-03-04

申请号：US13384990

申请日：2010-07-23

申请人： Olof Ericsson ,  Magnus Isaksson ,  Henrik Johansson ,  Ulf Landegren

发明人： Olof Ericsson ,  Magnus Isaksson ,  Henrik Johansson ,  Ulf Landegren

IPC分类号： C12Q1/68 ,  C40B30/04 ,  C12N15/09 ,  C12Q1/44

CPC分类号： C12Q1/6855

摘要： The present invention provides a method for detecting or enriching for a target deoxyribonucleic acid (DNA) present in a nucleic acid sample, said method comprising: (a) fragmenting a nucleic acid sample to generate nucleic acid fragments including a target fragment containing said target DNA and non-specifically ligating an adaptor sequence to an end of said fragments; (b) rendering said fragments at least partially single-stranded; (c) contacting the at least partially single-stranded fragments of step (b) with oligonucleotides A and B of a single target-specific nucleic acid probe; (d) ligating oligonucleotide B of said probe to the part of the single-stranded portion of said target fragment which is hybridised to oligonucleotide A of said probe to produce a probe-target fragment hybrid; and (e) detecting or enriching for said probe-target fragment hybrid.

摘要翻译： 本发明提供了用于检测或富集存在于核酸样品中的靶脱氧核糖核酸（DNA）的方法，所述方法包括：（a）将核酸样品片段化以产生包含含有所述靶DNA的靶片段的核酸片段 并且非特异性地将衔接子序列连接到所述片段的末端; （b）使所述片段至少部分单链; （c）使步骤（b）的至少部分单链片段与单个靶特异性核酸探针的寡核苷酸A和B接触; （d）将所述探针的寡核苷酸B连接到与所述探针的寡核苷酸A杂交的所述靶片段的单链部分的部分，以产生探针 - 靶片段杂交体; 和（e）检测或富集所述探针 - 靶片段杂交体。

公开(公告)号：US20100291636A1

公开(公告)日：2010-11-18

申请号：US12664177

申请日：2008-06-11

申请人： Henrik Johansson ,  Olof Ericsson

发明人： Henrik Johansson ,  Olof Ericsson

IPC分类号： C12P19/34

CPC分类号： C12N15/1065 ,  C12N15/10

摘要： The invention relates to a method for introducing common and/or individual sequence elements in a target nucleic acid molecule in a sample containing sample nucleic acid molecules, comprising the steps: i) denaturing the sample nucleic acid molecules, if the sample nucleic acid molecules are double-stranded, to obtain single stranded sample nucleic acid molecules; ii) bringing the sample nucleic acid molecules in contact with primary, secondary and tertiary probe nucleic acid molecules, wherein the 3′-end of the tertiary probe comprise a part complementary to the primary probe and the 5′-end of the tertiary probe comprise a part complementary to a 5′-part of the target nucleic acid molecule; the 3′-end of the secondary probe is complementary to a 3′-part of the target nucleic acid molecule and the 5′-end of the secondary probe is not complementary to the target nucleic acid molecule; wherein said primary, secondary and tertiary probes comprise said common and/or individual sequence elements; iii) ligating the 3′-end of the primary probe to the 5′-end of the target nucleic acid molecule; and iv) elongating the 3′-end of the secondary probe by means of a nucleic acid polymerase; or iv′) elongating the 3′-end of the target nucleic acid molecule.

摘要翻译： 本发明涉及在含有样本核酸分子的样品中引入目标核酸分子中的常见和/或单个序列元件的方法，其包括以下步骤：i）如果样品核酸分子为 双链，获得单链样品核酸分子; ii）使样品核酸分子与初级，次级和第三级探针核酸分子接触，其中第三级探针的3'-末端包含与主要探针互补的部分，并且第三探针的5'-末端包含 与靶核酸分子的5'-部分互补的部分; 第二探针的3'-末端与靶核酸分子的3'-部分互补，并且第二探针的5'-末端与靶核酸分子不互补; 其中所述初级，次级和第三级探针包括所述共同和/或单个序列元件; iii）将主要探针的3'-末端连接到靶核酸分子的5'-末端; 和iv）通过核酸聚合酶延伸第二探针的3'-末端; 或iv'）延伸靶核酸分子的3'末端。