CN1617937A - 标记过的核苷酸 - Google Patents

标记过的核苷酸 Download PDF

Info

Publication number
CN1617937A
CN1617937A CNA028278321A CN02827832A CN1617937A CN 1617937 A CN1617937 A CN 1617937A CN A028278321 A CNA028278321 A CN A028278321A CN 02827832 A CN02827832 A CN 02827832A CN 1617937 A CN1617937 A CN 1617937A
Authority
CN
China
Prior art keywords
nucleotide
mark
remove
joint
molecule
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CNA028278321A
Other languages
English (en)
Inventor
S·巴拉苏布拉马尼安
C·巴恩斯
刘小海
H·斯维尔德罗
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Solexa Ltd Great Britain
Original Assignee
Solexa Ltd Great Britain
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Family has litigation
First worldwide family litigation filed litigation Critical https://patents.darts-ip.com/?family=26246831&utm_source=google_patent&utm_medium=platform_link&utm_campaign=public_patent_search&patent=CN1617937(A) "Global patent litigation dataset” by Darts-ip is licensed under a Creative Commons Attribution 4.0 International License.
Priority claimed from GBGB0129012.1A external-priority patent/GB0129012D0/en
Application filed by Solexa Ltd Great Britain filed Critical Solexa Ltd Great Britain
Publication of CN1617937A publication Critical patent/CN1617937A/zh
Pending legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
    • C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
    • C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
    • C07H19/06—Pyrimidine radicals
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
    • C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
    • C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
    • C07H19/06—Pyrimidine radicals
    • C07H19/10—Pyrimidine radicals with the saccharide radical esterified by phosphoric or polyphosphoric acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
    • C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
    • C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
    • C07H19/16—Purine radicals
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
    • C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
    • C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
    • C07H19/16—Purine radicals
    • C07H19/20—Purine radicals with the saccharide radical esterified by phosphoric or polyphosphoric acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6869—Methods for sequencing
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2525/00—Reactions involving modified oligonucleotides, nucleic acids, or nucleotides
    • C12Q2525/10—Modifications characterised by
    • C12Q2525/186—Modifications characterised by incorporating a non-extendable or blocking moiety
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2535/00—Reactions characterised by the assay type for determining the identity of a nucleotide base or a sequence of oligonucleotides
    • C12Q2535/113—Cycle sequencing

Abstract

披露了通过可裂解的接头基团与可检测的标记连接的核苷和核苷酸。

Description

标记过的核苷酸
发明领域
本发明涉及标记过的核苷酸。具体地讲，本发明披露了具有可除去的标记的核苷酸，及其在多核苷酸测序方法中的用途。
背景
分子研究方面的进展部分是由于用于表征所述分子或它们的生物学反应的技术的改良。具体地讲，核酸DNA和RNA的研究，业已从用于序列分析的发展中的技术和杂交事件的研究中受益。
业已改善了核酸研究的所述技术的一种例子，是固定化核酸的组装的阵列的发展。所述阵列通常包括固定在固体支持材料上的多核苷酸的高密度距阵。例如，参见Fodor等，Trends Biotech.12：19-26，1994，该文献披露了组装核酸的方法，其中使用用掩膜保护的化学敏化的玻璃表面，不过，暴露了特定的部位，以便能够结合适当修饰过的核苷酸亚磷酰胺。组装的阵列还可以通过将已知的多核苷酸“点”在固体支持物的预定位置上而生产(例如Stimpson等，Proc.Natl.Acad.Sci.USA 92：6379-6383，1995)。
阵列技术的另一种发展，是将所述多核苷酸连接在固体支持材料上，以便形成单分子阵列。这种类型的阵列披露于国际专利申请号WO00/06770中。所述阵列的优点是，可以在单分子水平上监测反应，并且可以从单个反应中采集到大量单分子的信息。
为了使DNA阵列能够使用，必须确定所述分子的序列。美国专利号5,302,509披露了对固定在固体支持物上的多核苷酸进行测序的方法。所述方法取决于在存在DNA聚合酶的条件下，具有不同荧光标记的3′-封闭的碱基A，G，C和T向所述固定化多核苷酸中的掺入。所述聚合酶能够掺入与所述靶核苷酸互补的碱基，但是，能防止进一步添加3′-封闭基团。然后，可以确定所述掺入碱基的标记，并且通过化学裂解除去所述封闭基团，以便可以发生进一步的聚合。
Welch等(Chem.Eur.J.5(3)：951-960，1999)披露了用3′-O-封闭基团修饰的核苷三磷酸的合成，所述封闭基团是光不稳定性的，并且能发出荧光。打算将所述修饰过的核苷酸用于DNA测序实验。不过，证明了所述核苷酸难于掺入在现有的多核苷酸上，因为它不能结合到所述聚合酶的活性位点。
Zhu等(Cytometry 28：206-211，1997)还披露了通过所述碱基基团连接在所述核苷酸上的荧光标记的用途。希望将所述标记过的核苷酸，用于在原位杂交(FISH)实验中发出荧光，其中，需要一系列掺入的标记过的核苷酸，以便产生荧光“条形码”。
发明概述
在本发明中，通过与碱基连接的可裂解的接头基团，将核苷或核苷酸分子连接在可检测的标记上，使得所述分子可用于使用标记过的核苷或核苷酸的技术中，例如，测序反应，多核苷酸合成，核酸扩增，核酸杂交分析，单核苷酸多态性研究，以及其他使用诸如聚合酶，逆转录酶，末端转移酶，或其他DNA修饰酶的酶的其他技术。本发明特别适用于使用标记过的dNTPs的技术，如切口翻译，随机引物标记，末端标记(例如，使用末端脱氧核苷酸转移酶)，逆转录或核酸扩增。本发明的分子与现有技术不同，其中，所述标记连接在核糖或脱氧核糖上，或者，其中，所述标记是通过不可裂解的接头连接的。
根据本发明的第一方面，核苷酸或核苷分子或它们的类似物具有通过可裂解的接头连接在可检测的标记上的碱基。
本发明涉及具有通过可裂解的接头连接在可检测标记上的碱基的核苷酸或核苷分子。所述碱基可以是嘌呤或嘧啶。所述碱基可以是脱氮嘌呤。所述分子可以具有核糖或脱氧核糖糖部分。所述核糖或脱氧核糖可以包括通过2’或3’氧原子连接的保护基团。可以将所述保护基团除去，以便暴露出3’-OH基团。所述分子可以是脱氧核糖核苷三磷酸。所述可检测的标记可以是荧光团。所述接头可以是酸不稳定性接头，光不稳定性接头，或可以包括二硫键。
本发明还涉及标记核酸分子的方法，其中，所述方法包括将核苷酸或核苷分子掺入到所述核酸分子上，其中，所述核苷酸或核苷分子具有通过可裂解的接头连接在可检测的标记上的碱基。所述掺入步骤可以通过末端转移酶，聚合酶或逆转录酶完成。所述碱基可以是嘌呤或嘧啶。所述碱基可以是脱氮嘌呤。所述核苷酸或核酸分子可以具有核糖或脱氧核糖糖部分。所述核糖或脱氧核糖可以包括通过2’或3’氧原子连接的保护基团。可以将所述保护基团除去，以便暴露出3’-OH基团。所述分子可以是脱氧核糖核苷三磷酸。所述可检测的标记可以是荧光团。所述接头可以是酸不稳定性接头，光不稳定性接头，或可以包括二硫键。所述可检测的标记和/或可裂解的接头可以具有足以阻止第二个核苷酸或核苷掺入到所述核酸分子上的大小。
在另一方面，本发明涉及用于测定靶单链多核苷酸序列的方法，其中，所述方法包括监测互补性核苷酸的依次掺入，其中，每一个所述核苷酸具有通过可裂解的接头与可检测的标记连接的碱基。并且，其中，每一个掺入的核苷酸的身份是通过检测与所述碱基连接的标记确定的，并且，随后除去所述标记。
本发明还涉及用于测定靶单链多核苷酸的序列的方法，其中，该方法包括：(a)提供核苷酸，其中，所述核苷酸具有通过可裂解的接头与可检测的标记连接的碱基，并且，其中，所述与每一种类型的核苷酸连接的可检测的标记，在检测时可以与用于标记其他类型的核苷酸的可检测标记区分开来；(b)将核苷酸掺入到所述靶单链多核苷酸的互补体上；(c)检测(b)的核苷酸的标记，以便确定掺入的核苷酸的类型；(d)除去(b)的核苷酸的标记；和(e)任选重复步骤(b)-(d)一次或多次，以便测定靶单链多核苷酸的序列。
在本文所披露的方法中，可以让每一个核苷酸依次与所述靶接触，在添加下一个核苷酸之前除去未掺入的核苷酸，其中，所述标记的检测和除去是在添加每一个核苷酸之后，或在添加所有四种核苷酸之后进行的。
在所述方法中，可以让所有核苷酸同时与所述靶接触，即让包括所有不同核苷酸的组合物与所述靶接触，并且在检测之前除去未掺入的核苷酸，并且随后除去所述标记。
所述方法可以包括第一个步骤和第二个步骤，其中，在第一个步骤中，让包括四种核苷酸中的两种的第一种组合物与所述靶接触，并且在检测之前除去未掺入的核苷酸，并且随后除去所述标记，并且，其中，在第二个步骤中，让包括第一种组合物中所不包括的两种核苷酸的第二种组合物与所述靶接触，并且在检测之前除去未掺入的核苷酸，并且随后除去所述标记，并且，其中，任选重复所述第一个步骤和第二个步骤一次或多次。
本文所披露的方法还可以包括第一个步骤和第二个步骤，其中，在第一个步骤中，让包括四种核苷酸中的一种的组合物与所述靶接触，并且在检测之前除去未掺入的核苷酸，并且随后除去所述标记，并且，其中，在第二个步骤中，让包括第一种组合物中所不包括的三种核苷酸的第二种组合物与所述靶接触，并且在检测之前除去未掺入的核苷酸，并且随后除去所述标记，并且，其中，任选重复所述第一个步骤和第二个步骤一次或多次。
本文所披露的方法还可以包括第一个步骤和第二个步骤，其中，在第一个步骤中，将包括四种核苷酸中的三种的组合物与所述靶接触，并且在检测之前除去未掺入的核苷酸，并且随后除去所述标记，并且，其中，在第二个步骤中，让包括第一种组合物中所不包括的核苷酸的组合物与所述靶接触，并且在检测之前除去未掺入的核苷酸，并且随后除去所述标记，并且，其中，任选重复所述第一个步骤和第二个步骤一次或多次。
在另一方面，本发明涉及一种试剂盒，其中，所述试剂盒包括：(a)各个核苷酸，其中，每一个核苷酸具有通过可裂解的接头与可检测的标记连接的碱基，并且，其中，与每一种核苷酸连接的所述可检测的标记，在检测时可以与用于标记其他三种核苷酸的可检测标记区分开来；和(b)它们的包装材料。所述试剂盒还可以包括酶和适合所述酶作用的缓冲液。
所述核苷酸/核苷适用于很多不同的基于DNA的方法中，包括DNA合成和DNA测序方法。
根据本发明的另一方面，用于确定靶多核苷酸序列的方法包括监测互补性核苷酸的依次掺入，其中，所述核苷酸包括通过可裂解的接头与所述核苷酸的碱基部分连接的可检测的标记，通过监测所述标记而检测掺入，并且将所述标记除去，以便能够进行进一步的核苷酸掺入。
附图说明
图1表示用于本发明的典型核苷酸结构。对于每一种结构来说，X可以是H，磷酸，二磷酸或三磷酸。R1和R2可以相同或不同，并且可以选自H，OH，或可以转化成OH的任何基团。
图2表示用于本发明的接头的结构，除了可以使用的所述接头的一般定义之外，包括(1)二硫接头和酸不稳定性接头，(2)二烷氧基苄基接头，(3)Sieber接头，(4)吲哚接头和(5)叔丁基Sieber接头。
图3表示用于本发明的某些功能性分子，包括某些可裂解的接头。在所述结构中，R1和R2可以相同或不同，并且可以是H，OH，或可以转化成OH的任何基团，包括羰基。R3表示独立地选自烷基，烷氧基，氨基，或卤素的一个或多个取代基。另外，可以用任何用在3′-封闭基团上的不稳定性官能团，构建可裂解的接头。
图4表示变性凝胶，展示用Klenow聚合酶进行的实施例1的三磷酸的掺入。
图5表示变性凝胶，展示用Klenow聚合酶进行的实施例3的三磷酸的掺入。
图6表示变性凝胶，展示用Klenow聚合酶进行的实施例4的三磷酸的掺入。
详细说明
本发明涉及通过可裂解的接头连接标记的方式修饰过的核苷酸和核苷，以便使得所述分子可用于所述标记过的分子与酶相互作用的技术中，如测序反应，多核苷酸合成，核酸扩增，核酸杂交分析，单核苷酸多态性研究，使用诸如聚合酶，逆转录酶，末端转移酶的酶的技术，使用标记过的dNTPs的技术(例如，切口翻译，随机引物标记，末端标记(例如，使用末端脱氧核苷酸转移酶)逆转录，或核酸扩增)。
正如本领域所公知的，“核苷酸”由含氮碱基，糖，以及一个或多个磷酸基团组成。在RNA中，所述糖是核糖，而在DNA中，所述糖是脱氧核糖，即缺少存在于核糖中的羟基的糖。所述含氮碱基是嘌呤或嘧啶的衍生物。所述嘌呤是腺嘌呤(A)和鸟嘌呤(G)，而所述嘧啶是胞嘧啶(C)和胸腺嘧啶(T)(或对于RNA来说，是尿嘧啶(U))。将脱氧核糖的C-1原子键合于嘧啶的N-1或嘌呤的N-9。核苷酸也是核苷的磷酸酯，酯化发生在与所述糖的C-5连接的羟基上。核苷酸通常是单磷酸，二磷酸或三磷酸酯。
“核苷”在结构上类似于核苷酸，但是，缺少了磷酸部分。核苷类似物的一种例子是其中的标记与所述碱基连接的核苷，并且，没有与糖分子结合的磷酸基团。
尽管所述碱基通常被称作嘌呤或嘧啶，技术人员可以理解的是，可以获得不会改变所述核苷酸或核苷进行Watson-Crick碱基配对的能力的衍生物和类似物。“衍生物”或“类似物”表示其核心结构与母体化合物相同或非常接近的化合物或分子，不过，它具有化学或物理修饰，如不同的或额外的侧基，这使得所述衍生的核苷酸或核苷能够与其他分子连接。例如，所述碱基可以是脱氮嘌呤。所述衍生物应当能够进行Watson-Crick配对。“衍生物”和“类似物”还表示具有修饰过的碱基部分和/或修饰过的糖部分的合成核苷酸或核苷衍生物。例如，所述衍生物和类似物披露于以下文献中：Scheit，NucleotideAnalogs(John Wiley & Son，1980)和Uhlman等，Chemical Reviews90：543-584，1990。核苷酸类似物还可以包括修饰过的磷酸二酯键，包括硫代磷酸酯，二硫代磷酸酯，烷基磷酸酯，phosphoranilidate和氨基磷酸酯键。所述类似物应当能够进行Watson-Crick碱基配对。本文所使用的“衍生物”和“类似物”可以交换使用，并且包括在本文所定义的术语“核苷酸”和“核苷”的范围内。
本发明可以利用常规可检测标记。检测可以通过任何合适的方法进行，包括荧光光谱法或通过其他光学方法。优选的标记是荧光团，它在吸收能量之后，能发射出特定波长的辐射。很多合适的荧光标记是已知的。例如，Welch等(Chem.Eur.J.5(3)：951-960，1999)披露了丹酰-官能化荧光部分，可将它用于本发明中。Zhu等(Cytometry28：206-211，1997)披露了荧光标记Cy3和Cy5的用途，还可将其用于本发明中。适合使用的标记还披露于以下文献中：Prober等(Science 238：336-341，1987)；Connell等(BioTechniques 5(4)：342-384，1987)，Ansorge等(Nucl.Acids Res.15(11)：4593-4602，1987)和Smith等(Nature 321：674，1986)。其他可通过商业渠道获得的荧光标记包括，但不局限于荧光素，罗丹明(包括TMR，德克萨斯红和Rox)，alexa，bodipy，吖啶，香豆素，芘，苯并蒽和花青苷。
还可以将多种标记用于本发明中。例如，双荧光团FRET盒(Tet.Letts.46：8867-8871，2000)为本领域所熟知，并且可用于本发明中。还可以使用多荧光树枝型聚合物系统(J.Amer.Chem.Soc.123：8101-8108，2001)。
尽管荧光标记是优选的，对于本领域普通技术人员来说，其他形式的可检测标记显然是可利用的。例如，微颗粒，包括量子斑点(Empodocles等，Nature 399：126-130，1999)，金纳米颗粒(Reichert等，Anal.Chem.72：6025-6029，2000)，微珠(Lacoste等，Proc.Natl.Acad.Sci USA 97(17)：9461-9466，2000)，以及可以通过质谱法检测的标记都可以使用。
还可以将多成分标记用于本发明中。多成分标记是取决于与用于检测的另一种化合物相互作用的标记。生物学中所使用的最常见的多成分标记是生物素-链亲和素系统。将生物素用作与所述核苷酸碱基连接的标记。然后单独添加链亲和素，以便能够进行检测。可以获得其他多成分系统。例如，二硝基苯酚具有可以通过商业渠道获得的能用于检测的荧光抗体。
所述标记(或标记和接头构建体)可以具有足以发挥阻断其他核苷酸掺入到本发明的核苷酸上的大小或结构。这使得能够进行受控制的聚合。所述阻断可能是由于空间位阻造成的，或者可能是由于大小，电荷和结构的组合造成的。
将针对核苷酸对本发明作进一步说明。不过，除非另有说明，对核苷酸的说明同样适用于核苷。还将针对DNA对本发明作进一步说明，不过，这种说明还适用于RNA，PNA，以及其他核酸，除非另有说明。
本发明的修饰过的核苷酸利用可裂解的接头将所述标记连接在核苷酸上。可裂解的接头的使用，能确保所述标记可以在检测之后除去(如果必要的话)，避免了任何干扰信号，并且随后可以掺入任何标记过的核苷酸。
可裂解的接头为本领域所公知，并且可以采用常规化学方法，以便将接头连接在核苷酸碱基和标记上。所述接头可以通过任何合适的方法裂解，包括接触酸，碱，亲核试剂，亲电试剂，自由基，金属，还原或氧化试剂，光照，温度，酶等。合适的接头可以用标准化学封闭基团改良，正如在以下文献中所披露的：Greene & Wuts，ProtectiveGroups in Organic Synthesis，John Wiley & Sons。Guillier等披露了用于固相合成的其他合适的可裂解的接头(Chem.Rev.100：2092-2157，2000)。
术语“可裂解的接头”的使用并非意味着需要将整个接头从核苷酸碱基上除去。裂解位点可能位于所述接头上的位置，以便确保在裂解之后所述接头的部分保持与所述核苷酸碱基连接。
所述接头可以连接在核苷酸碱基的任何位置上，其前提是，仍然能进行Watson-Crick碱基配对。对于嘌呤碱基来说，如果所述接头是通过所述嘌呤或优选的脱氮嘌呤类似物的7号位置，通过8-修饰过的嘌呤，通过N-6修饰过的腺嘌呤或N-2修饰过的鸟嘌呤连接的话将是优选的。对于嘧啶来说，连接优选是通过胞嘧啶，胸腺嘧啶和尿嘧啶上的5号位置和胞苷上的N-4位置连接的。合适的核苷酸结构如图1所示。对于图1中的每一种结构来说，X可以是H，磷酸，二磷酸或三磷酸。R1和R2可以相同或不同，并且可以选自H，OH，或能够转化成OH的任何基团，包括，但不局限于羰基。
合适的接头总体上在图2中示出，并且，包括，但不局限于二硫接头(1)，酸不稳定性接头(2，3，4和5；包括二烷氧基苄基接头(例如2)，Sieber接头(例如)，吲哚接头(例如4)，叔丁基Sieber接头(例如))，亲电可裂解的接头，亲核可裂解的接头，光可裂解的接头，在还原条件，氧化条件下裂解，通过使用保险栓(safety-catch)接头进行的裂解，以及通过消除机制进行裂解。
A.亲电子裂解的接头。
亲电子裂解的接头通常是由质子裂解的，并且包括对酸的裂解敏感性。合适的接头包括修饰过的苄基系统，如三苯甲基，对烷氧基苄基酯，和对烷氧基苄基酰胺。其他合适的接头包括叔丁氧羰基(Boc)基团，和缩醛系统(例如，在图3中示出的O-C(R4)(R5)-O-R6)。
为了制备合适的接头分子，还可以考虑将诸如镍，银或汞的亲硫金属用于硫代缩醛或其他含硫的保护基团的裂解。
B.亲核裂解的接头
亲核裂解也是得到充分认可制备接头分子的的方法。可以使用在水中不稳定的诸如酯的基团(即，可以在碱性pH下简单地裂解)，以及对非水亲核试剂不稳定的基团。可以用氟离子裂解诸如三异丙基硅烷(TIPS)或叔丁基二甲基硅烷(TBDMS)的基团中的硅-氧键。
C.光可裂解的接头。
光可裂解的接头业已被广泛用于糖类化学。优选的是，激活裂解所需要的光不会影响所述修饰过的核苷酸的其他成分。例如，如果用荧光团作标记的话，如果它所吸收的光的波长与裂解所述接头分子所需要的光的波长不同，则是优选的。合适的接头包括基于O-硝基苄基化合物和硝基藜芦基化合物的接头。还可以使用基于二苯乙醇酮化学的接头(Lee等，J.Org.Chem.64：3454-3460，1999)。
D.在还原条件下的裂解
存在很多已知容易受还原性裂解的接头。业已将使用基于钯的催化剂的催化氢化作用裂解苄基和苄基氧基羰基基团。二硫键还原也是本领域所公知的。
E.在氧化条件下的裂解
基于氧化的方法为本领域所熟知。其中包括对烷氧基苄基的氧化以及硫和硒接头的氧化。利用含水的碘裂解二硫接头和其他基于硫或硒的接头也属于本发明的范围。
F.保险栓接头
保险栓接头是分两个步骤裂解的接头。在一种优选系统中，第一个步骤是制备反应性亲核中心，然后进行的第二个步骤包括会导致裂解的分子内环化。例如，可以用肼或光化学处理乙酰丙酸酯键，以便释放活性胺，然后可以将所述活性胺环化，以便裂解所述分子中其他部位的酯(Burgess等，J.Org.Chem.62：5165-5168，1997)。
G.通过消除机制裂解
还可以使用消除反应。例如，可以使用诸如Fmoc和氰乙基的基团的碱催化的消除，和烯丙基系统的钯催化的还原型消除。
除了所述裂解位点之外，所述接头可以包括间隔单位。所述间隔基团将所述核苷酸碱基与所述裂解位点或标记分开。所述接头的长度是不重要的，其前提是，所述标记保持离开所述核苷酸的足够距离，以便不干扰所述核苷酸和酶的任何相互作用。
所述修饰过的核苷酸还可以包括其他基团或所述糖基团的修饰。例如，可以制备在核糖环结构上缺少两个氧的双脱氧核糖衍生物(位于2′和3′位置上)，并且用作生长的寡核苷酸链上的其他核苷酸掺入的封闭基团。所述保护基团要被用于阻止核苷酸掺入在新生的多核苷酸链上，并且可以在特定条件下除去，以便能够进行聚合。与现有技术相反，没有结合在核糖3’位置上的可检测的标记。由此确保降低对所述聚合酶的空间位阻，但仍然能够用所述保护基团控制掺入。
技术人员将会理解如何将合适的保护基团连接在核糖环上，以便阻断与3′-OH的相互作用。所述保护基团可以直接连接在3’位置上，或者可以连接在2’位置上(所述保护基团具有足够的大小或电荷，以便阻断3’位置上的相互作用)。另外，所述保护基团可以连接在3′和2′位置，并且可以裂解，以便暴露出3′OH基团。
合适的保护基团对本发明技术人员来说是显而易见的，并且可以用披露于以下文献中的任何合适的保护基团制备：Green和Wuts，同上。所述保护基团应当是可以除去的(或可以修饰的)，以便产生3′OH基团。用于获得3′OH的方法，可以是任何合适的化学或酶促反应。
所述不稳定性的接头，可以包括能够在与所述封闭相同的条件下裂解的官能团。这将使得所述脱保护方法更为有效，因为裂解所述标记和封闭基团只需要一次处理。因此，所述接头可以包括如图3所示的官能团，它可以与位于所述残余核苷上的羟基官能团或所述除去的标记一起裂解。所述接头还可以包括完全不同的化学官能团，所述官能团对于用于裂解所述封闭基团的条件来说是不稳定的。
术语“烷基”包括直链和支链烷基，除非本文中另有说明，术语“烷基”表示具有1-8个碳原子的基团，并且通常具有1-6个碳原子，例如1-4个碳原子。烷基的例子，包括甲基，乙基，丙基，异丙基，正丁基，异丁基，叔丁基，正戊基，2-戊基，3-戊基，2-甲基丁基，3-甲基丁基和正己基以及它们的异构体。
环烷基的例子是具有3-10个环原子的基团，具体例子包括源于环丙烷，环丁烷，环戊烷，环己烷和环庚烷，双环庚烷和萘烷的基团。
链烯基的例子包括，但不局限于乙烯基(烯基)，1-丙烯基，2-丙烯基(烯丙基)，异丙烯基，丁烯基，丁-1，4-二烯基，戊烯基，和己烯基。
环链烯基的例子包括，但不局限于环丙烯基，环丁烯基，环戊烯基，环戊二烯基，和环己烯基。
术语“烷氧基”表示C1-6烷氧基，除非另有说明：-OR，其中，R是C1-6烷基。C1-6烷氧基的例子包括，但不局限于-OMe(甲氧基)，-OEt(乙氧基)，-O(nPr)(正丙氧基)，-O(iPr)(异丙氧基)，-O(nBu)(正丁氧基)，-O(sBu)(仲-丁氧基)，-O(iBu)(异丁氧基)，和-O(tBu)(叔-丁氧基)。
术语氨基表示NR1R2类型的基团，其中，R1和R2独立地选自氢，C1-6烷基(又称之为C1-6烷基氨基或二-C1-6烷基氨基)。
本文所使用的术语“卤素”包括氟，氯，溴，和碘。
本发明的核苷酸分子适用于很多不同的方法，其中，需要检测核苷酸。
DNA测序方法，如可以用所述核苷酸进行披露于美国专利号5,302,509中的方法。
用于测定靶多核苷酸序列的方法可以这样进行：让靶多核苷酸分别与不同的核苷酸接触，以便形成所述靶核苷酸的互补体，并且检测所述核苷酸的掺入。所述方法利用了聚合，使得聚合酶通过掺入互补于所述靶的正确的核苷酸，以延伸所述互补链。所述聚合反应还需要特殊引物来启动聚合作用。
对每一轮反应来说，所述标记过的核苷酸的掺入，是通过聚合酶进行的，并且，然后测定所述掺入事件。存在很多不同的聚合酶，并且，对本领域普通技术人员来说，显而易见的是哪一种酶是最适合使用的。优选的酶包括DNA聚合酶I，Klenow片段，DNA聚合酶III，T4或T7 DNA聚合酶，Taq聚合酶或vent聚合酶。还可以使用通过工程方法改造成具有特定性质的聚合酶。
所述测序方法优选对排列在固体支持物上的靶多核苷酸进行。可以通过接头分子将多个靶多核苷酸固定在所述固体支持物上，或者可以连接在诸如微球体的颗粒上，所述颗粒还可以连接在固体支持材料上。
可以通过多种方法将所述多核苷酸连接在所述固体支持物上，包括使用生物素-链亲和素相互作用。用于将多核苷酸固定在固体支持物上的方法为本领域所公知，并且，包括石板印刷技术以及将每一种多核苷酸“点”在固体支持物的特定位置上。合适的固体支持物为本领域所公知，并且包括玻璃载玻片和珠，陶瓷和硅表面和塑料材料。所述支持物通常是平面，尽管也可以使用微珠(微球体)，并且还可以通过已知方法将后者连接在其他固体支持物上。所述微球体可以具有任何合适的大小，其直径通常为10-100纳米。在优选实施方案中，将所述多核苷酸直接连接在平面上，优选连接在平的玻璃表面上。连接优选通过共价键的形式进行。所使用的阵列优选是单分子阵列，它包括位于独特的光学可分辨区域的多核苷酸，例如，正如在国际申请号WO00/06770中所披露的。
所述测序列方法可以针对单个多核苷酸分子和多个多核苷酸分子阵列进行，即具有不同的各个多核苷酸分子的阵列，和具有包括一种多核苷酸分子的多个拷贝的不同区域的阵列。单分子阵列使得每一种多核苷酸可以独立分辨。优选使用单分子阵列。对单一分子阵列的非破坏性测序，使得能够形成可以空间寻址的阵列。
所述方法利用聚合反应制备所述靶的互补序列。进行聚合的必须条件，对技术人员来说是显而易见的。
为了进行所述聚合酶反应，通常首先必须使引物序列与所述靶多核苷酸退火，所述引物序列是由所述聚合酶识别的，并且起着所述互补链随后延伸的起始位点的作用。所述引物序列可以相对所述靶多核苷酸作为独立的成分添加。另外，所述引物和靶多核苷酸可以分别是一个单链分子的一部分，由所述引物部分与所述靶的一部分形成分子内双链体，即发卡环结构。可以在所述分子的任何位点，将该结构固定在所述固体支持物上。进行所述聚合酶反应所必需的其他条件，对本领域技术人员来说是显而易见的，这些条件包括温度，pH，缓冲液组成。
然后，让本发明的修饰过的核苷酸与所述靶多核苷酸接触，以便能够进行聚合。所述核苷酸可以依次添加，即分别添加每一种类型的核苷酸(A，T，G或C)，或同时添加。如果所述核苷酸是同时添加的话，优选用不同的标记对每一种类型的核苷酸进行标记。
让所述聚合步骤进行足以掺入一个核苷酸的时间。
然后除去未掺入的核苷酸，例如，通过对所述阵列实施洗涤步骤，并且随后可以进行对掺入标记的检测。
检测可以通过常规方法进行，例如，如果所述标记是荧光部分的话，掺入的碱基的检测，可以通过使用共焦扫描显微镜进行，用激光扫描所述阵列的表面，以便使直接结合在所述掺入的碱基上的荧光团成像。另外，可以用灵敏的2-D探测器，如电荷偶连的探测器(CCD)观察所产生的每一种信号。不过，可以获得诸如扫描近场光学显微方法(SNOM)的其他技术，并且可用于使密集的阵列成像。例如，使用SNOM可以分辨间隔距离小于100纳米，例如，10纳米-10微米的各个多核苷酸。有关扫描近场光学显微方法的说明，可以参见Moyer等，Laser Focus World 29：10，1993。用于使多核苷酸阵列成像的合适的装置是公知的，并且其技术参数为技术人员所公知。
在检测之后，可以用裂解所述接头的合适条件除去所述标记。
修饰过的核苷酸的使用并不局限于DNA测序技术，还可用本发明的核苷酸实施包括多核苷酸合成，DNA杂交分析，和单核苷酸多态性研究的其他形式。涉及到核苷酸和酶之间的相互作用的任何技术，都可以利用本发明的分子。例如，可以将所述分子用作逆转录酶或末端转移酶的底物。
合适的结构披露于以下实施例中，并且在附图中示出。
实施例
实施例1.二硫接头的合成
Figure A0282783200181
将叔丁基-N-(2-巯基乙基)氨甲基酸酯(3mmol，0.5mL)滴加到将1.32g(6.0mmol)aldrithiol溶解在15mL MeOH中制成的溶液中。在1.5小时之后，该反应业已结束，并且蒸发所述溶剂。通过在二氧化硅上面用乙酸乙酯∶石油醚(1∶4)层析而纯化所述粗制产物。产物1a是以浅黄色油状物形式获得的(0.76g，2.67mmol，89％)。1HNMR(500Mhz，D6-DMSO)：d＝1.38(s，9H，tBu)，2.88(t，J＝6.6Hz，2H，SCH2)，3.20(q，J＝6.6Hz，2H，CH2NH)，7.02(bs，1H，NH)，7.24(ddd，J＝7.3Hz，J＝4.9Hz，J＝1.0Hz，1H，H-5)，7.77(dt，J＝8.1Hz，J＝1.0Hz，1H，H-3)，7.82(ddd，J＝8.1Hz，J＝7.4Hz，J＝1.8Hz，1H，H-4)，8.46(ddd，J＝4.9Hz，J＝1.8Hz，J＝1.0Hz，1H，H-6)。
Figure A0282783200182
为了对1a的胺脱保护，将17mg(60μmol)溶解在0.5mL DCM和0.5mL三氟乙酸的混合物中。在室温下将该混合物搅拌2.5小时，然后在减压条件下除去溶剂。重复三次将残余物重新溶解在2mL DCM中，并且蒸发至干燥。在高真空条件下将所述脱保护的产物干燥3小时，然后溶解在1mL干燥DMF中。假设所述脱保护作用业已完成。
向将15mg 5-羧甲基四甲基罗丹明(35μmol)溶解在2mL DMF中制成的溶液中添加8.0mg N-羟基琥珀酰亚胺(70μmol)和7.8mg DCC(38μmol)。在黑暗中搅拌该混合物6小时。然后添加22μl DIPEA(126μmol)和溶解在1mL DMF中的脱保护的1a的溶液。在黑暗中搅拌该反应混合物过夜，然后在减压条件下除去溶剂。将残余物溶解在DCM中，并且用饱和氯化钠溶液洗涤。在硫酸镁上干燥之后，用CHCl3∶MeOH(3∶1)作溶剂在二氧化硅上纯化所述粗制混合物。1b是以暗红色固体形式分离的，产率为90％(19.2mg，31.4μmol)。1H NMR(500MHz，D6-DMSO)：δ＝3.09(t，J＝6.7Hz，2H，SCH2)，3.63(q，J＝6.2Hz，2H，CH2NH)，6.48-6.53(m，6H，H-蒽)，7.23-7.26[m，1H，(吡啶)]，7.32(d，J＝7.9Hz，1Hz，H-3)，7.81-7.82[m，2H，H-3+H-4(吡啶)]，8.21(d，J＝7.9Hz，1H，H-4)，8.43(s，1H，H-6)，8.47[dt，J＝4.7Hz，J＝1.3Hz，1H，H-6(吡啶)]，9.03(t，J＝5.2Hz，1H，NH)。
将巯基丙酸(20.6μmol，1.8ml)添加到由19.6mg 1b(32.7μmol)溶解在2mL MeOH中制成的溶液中。在黑暗中搅拌2.5小时。在减压条件下除去所述溶剂。用CHCl3∶MeOH∶AcOH 15∶1∶0.5作溶剂混合物，通过在二氧化硅上层析，纯化所述粗制产物。可以分离15.5mg(26μmol，80％)暗红色晶体1c。1H NMR(500MHz，D20)：δ＝2.53(t，J＝7.0Hz，2H，CH2COOH)，2.88(t，J＝7.0Hz，2H，CH2CH2COOH)，2.96-2.99(m，2H，CH2CH2NH)，3.73(t，J＝6.3Hz，2H，CH2NH)，6.53(d，J＝2.4Hz，2H，H-蒽)，6.81(dd，J＝9.5Hz，J＝4.5Hz，2H，H-蒽)，7.12(d，J＝9.5Hz，2H，H-蒽)，7.48(d，J＝7.9Hz，1H，H-3)，7.95(dd，J＝8.1Hz，J＝1.9Hz，1H，H-2)，8.13(d，J＝1.9Hz，1H，H-1)。+v电子喷射(C30H31N3O6S2)：预计为593.17；实测为594.3[M+H]，616.2[M+Na]。
将9.9mg N-羟基琥珀酰亚胺(86.8μmol)和9.7mg DCC(47.1μmol)添加到通过将25.8mg 1c(43.4μmol)溶解在3mL DMF(无水)制成的溶液中。在室温下，在黑暗中将该混合物搅拌5小时，然后放入冰箱中过夜。通过棉绒塞将该混合物过滤到新的烧瓶中，并且向该混合物中添加865μL propargylamino dUTP(14.7μmol，17μmol，溶解在1mL H2O)和3mL硼酸钠缓冲液(0.1M溶液，pH9)的溶液中。搅拌该混合物过夜。在除去溶剂之后，将残余物溶解在尽可能少的水中，并且通过HPLC纯化。使用Zorba×C18柱，用0.1M三乙基碳酸氢铵(TEAB)和乙腈作缓冲液。31p NMR(400MHz，D2O)：δ＝-4.73(d)，-9.93(d)，19.03(t)。-ve电子喷射(C42H47N6O19P3S2假定4H+抗衡离子)。预计为1096.16；实测为1092.9。在水中的UV：λ(max)＝555nm，A(555)＝0.885(c＝0.036μmol)。
用Klenow DNA聚合酶成功地掺入了三磷酸(1)。所述反应是在以下条件下进行的：50mM Tris·HCl(pH7.5)，10mM NaCl，2mM DTT，0.1mM EDTA，5mM MgCl2，2μM化合物3，100nM DNA模板(事先用P32和T4多核苷酸激酶标记过的)和10个单位的市售外-Klenow(Amersham Corp.，Arlington Heights，Illinois，USA)。所述DNA模板是自我互补的发卡结构(5′-TACCgTCgACgTCgACgCTggCg-AgCgTgCTgCggTTTTT(C6-氨基)TTACCgCAgCACgCTCgCCAgCg；SEQ ID NO：1)。所述反应是在37℃下，在100μL体积中进行的，时间点选择为0，1，3，5和10分钟。使所述反应产物沿变性的(8M尿素)20％聚丙烯酰胺凝胶电泳，并且在typhoon磷成像仪上成像。在1分钟内观察到了完全的单碱基延伸，表明具有有效的聚合酶掺入(二硫接头凝胶，图4)。示出了第二组泳道，其中，在掺入之后，让所述材料接触DTT。可以看到一个不同的带移动，表明从所述DNA构建体上除去了所述染料。因此，业已使用这种二硫化物化合物证实了一轮聚合酶掺入和裂解。
实施例2.无TMR-Sieber接头的酸的合成。
将5-[-9-[9-(芴基-甲基氧基羰基)氨基]呫吨-3-基]戊酸(42.8mg，80μmol)在室温下，与二琥珀酰碳酸酯(22.5mg，88μmol)和N，N-二甲基氨基吡啶(10.8mg，88μmol)一起在DMF中搅拌。5分钟之后，添加单-5-羧基TMR乙二胺(198.9mg，40μmol)，然后添加DIPEA(13.9μl，80μmol)。在室温下搅拌所述反应混合物。2小时之后，用二氯甲烷(100mL)稀释所述反应混合物，并且用1M含水的磷酸二氢钾(50mL)提取所得到的溶液。分离DCM层，并且在减压下蒸发。通过短的柱层析纯化所述残余物。收集用在氯仿中配制的40％甲醇洗脱的所述级份，并且在减压条件下蒸发。然后，将所述残余物溶解在无水DMF(1mL)和N-(2-巯基乙基)氨基甲基聚苯乙烯(200mg，400μmol)和DBU(12μL，80μmol)中。在室温下放置10分钟之后，过滤所述残余物，并且用无水DMF(1mL)漂洗。合并所有的滤液，然后添加到将琥珀酸酐(80mg，800μmol)，DIPEA(139μL，800μmol)和DMAP(9.8mg，80μmol)溶解在DMF(1mL)中制备的溶液中。然后，在室温下搅拌所述反应混合物。在过夜(16小时)之后，在减压下蒸发所有的溶剂，并且通过短的柱层析纯化所述残余物。以紫色粉末形式获得了用溶解在氯仿中的30％的甲醇洗脱的标题化合物(22mg，总产率63％)。1HNMR[D6-DMSO]：8.82(1H，t，J5.4，ex.)，8.75(1H，d，J8.9，ex.)，8.42(1H，d，J1.5)，8.20(1H，dd，J8.0和1.5)，7.95(1H，t，J5.9，ex.)，7.34(1H，d，J7.3)，7.30-7.27(2H，m)，7.21(1H，d，J8.5)，7.16-7.07(2H，m)，6.68(1H，dd，J8.8和2.5)，6.65(1H，d，J2.4)，6.49-6.43(6H，m)，6.18(1H，d，J5.6)，3.95(1H，t，J5.9)，3.39-3.36(2H，m)，3.30-3.27(2H，m)，2.92(12H，s)，2.37-2.33(2H，m)，2.14(2H，t，J7.2)和1.70-1.62(4H，m)。MS[(ES(+)]，m/z 868.5(MH+)。
实施例3.TMR-Sieber接头-dUTP(3)的合成
Figure A0282783200221
在室温下，将无TMR-sieber接头的酸(4.34mg，5μmol)与二琥珀酰碳酸酯(1.74mg，7.5μmol)和N，N-二甲基氨基吡啶(0.92mg，7.5μmol)在DMF(1mL)中搅拌。10分钟之后，将所有反应温和物添加到5-(3-氨基丙炔基)-2′-脱氧尿苷-5′-三磷酸的四-(三-丁铵)盐(10mL)中。在室温下搅拌该反应混合物4小时，并且在冰箱中保存过夜。然后用冷却水(10mL)稀释所述反应混合物，并且将所有得到的溶液加样到DEAE A-25的短柱上。首先用0.1M TEAB缓冲液洗脱所述柱，然后用0.7M TEAB缓冲液洗脱。收集0.7M TEAB洗脱剂，并且在减压下蒸发。用MeOH(2×10mL)共同蒸发残余物，然后通过制备HPLC纯化。以三乙铵盐的形式获得了标题化合物，产率为31％(根据在水中，在555nm波长下对TMR的定量测定(pH7))。在D2O中的1HNMR表明了两种非对映异构体，这是由于sieber接头部分，并且存在大约三个三乙铵抗衡离子。1HNMR[D2O]：8.18(1H，m)，8.06(1H，m)，7.76(0.55H，s)，7.74(0.45H，s)，7.36-7.09(5H，m)，6.89-6.72(3H，m)，6.59-6.37(5H，m)，6.12(0.55H，t，J6.6)，6.05(0.45H，t，J6.6)，5.99(0.45H，d，J2.5)，5.91(1.1H，m)，5.88(0.45H，s)，4.49(0.55H，m)，4.43(0.45H，m)，4.00-3.35(9H，m)，3.30-2.95(32H，m)，2.65-2.52(4H，m)，2.25-2.05(4H，m)，1.62-1.42(4H，m)和1.23(27H，t，J7.3)。31P[D2O]：-9.91(γP，d，J19.2)，[-11.08(αP，d，J20.1)，和-11.30(αP，d，J20.1)，由于两种非对称异构体1和-22.57(βP，m)。MS[(ES(-))m/z1369.1(M-)。
用Klenow DNA聚合酶成功地掺入了三磷酸(3)。所述反应是在以下条件下进行的：50mM Tris-HCl(pH7.5)，10mM NaCl，2mM DTT，0.1mM EDTA，5mM MgCl2，2μM化合物3，100nM DNA模板(事先用P32和T4多核苷酸激酶标记过)和10个单位的市售外-Klenow(Amersham Corp.Arlington Heights，Illinois，USA)。所述DNA模板是自我互补的发卡结构(5′-TACCgTCgACgTCgACgCTggCg-AgCgTgCTgCggTTTTT(C6-氨基)TTACCgCAgCACgCTCgCCAgCg；SEQ ID NO：1)。所述反应是在37℃下，在100μL体积中进行的，时间点选择为0，1，3，5和10分钟。使所述反应产物沿变性的(8M尿素)20％聚丙烯酰胺凝胶电泳，并且在typhoon磷成像仪上成像。在1分钟内出现了完全的单碱基延伸,表明有效的聚合酶掺入(Sieber接头凝胶，图5)。
实施例4.TMR-吲哚接头-dUTP(4)的合成
Figure A0282783200241
用KlenowDNA聚合酶成功地掺入了三磷酸(4)。所述反应是在以下条件下进行的：50mM Tris·HCl(pH7.5)，10mM NaCl，2mM DTT，0.1mM EDTA，5mM MgCl2，2μM化合物3，100nM DNA模板(事先用P32和T4多核苷酸激酶标记过)和10个单位的市售外-Klenow(Amersham Corp.Arlington Heights，Illinois，USA)。所述DNA模板是自我互补的发卡结构(5′-TACCgTCgACgTCgACgCTggCg-AgCgTgCTgCggTTTTT(C6-氨基)TTACCgCAgCACgCTCgCCAgCg；SEQ ID NO：1)。所述反应是在37℃下，在100μL体积中进行的，时间点选择为0，1，3，5和10分钟。使所述反应产物沿变性的(8M尿素)20％聚丙烯酰胺凝胶电泳，并且在typhoon磷成像仪上成像。在1分钟内观察到了完全的单碱基延伸，表明有效的聚合酶掺入(吲哚接头凝胶，图6)。
本文所引用的所有专利，专利申请，和公开的参考文献被以它们的全文形式收作本文参考。尽管业已结合本发明的优选实施方案对本发明进行了具体图示和说明，本领域技术人员可以理解的是，可以对其中的形式和细节进行各种改变，而又不超出由所附权利要求书所包含的本发明的范围。

Claims (25)

1.一种核苷酸或核苷分子，具有通过可裂解的接头与可检测的标记连接的碱基。
2.如权利要求1的分子，其中，所述碱基是嘌呤或嘧啶。
3.如权利要求1或2的分子，其中，所述碱基是脱氮嘌呤。
4.如权利要求1-3中任意一项的分子，具有核糖或脱氧核糖糖部分。
5.如权利要求4的分子，其中，所述核糖或脱氧核糖包括通过2′或3′氧原子连接的保护基团。
6.如权利要求1-5中任意一项的分子，它是脱氧核糖核苷三磷酸。
7.如权利要求1-6中任意一项的分子，其中，所述可检测的标记是荧光团。
8.如权利要求1-7中任意一项的分子，其中，所述接头是酸不稳定性的，光不稳定性的，或包括二硫键。
9.一种标记核酸分子的方法，该方法包括将核苷酸或核苷分子掺入到所述核酸分子中，其中，所述核苷酸或核苷分子具有通过可裂解的接头与可检测的标记连接的碱基。
10.如权利要求9的方法，其中，所述掺入是通过末端转移酶，聚合酶或逆转录酶进行的。
11.如权利要求9或10的方法，其中，所述碱基是脱氮嘌呤。
12.如权利要求9-11中任意一项的方法，其中，所述核苷酸或核苷分子具有核糖或脱氧核糖糖部分，并且，其中，所述核糖或脱氧核糖包括通过2′或3′氧原子连接的保护基团，并且可以对它进行修饰或将其除去，以便暴露出3′OH基团。
13.如权利要求9-12中任意一项的方法，其中，所述核苷酸是脱氧核糖核苷酸三磷酸。
14.如权利要求9-13中任意一项的方法，其中，所述标记是荧光团。
15.如权利要求9-14中任意一项的方法，其中，所述接头是酸不稳定性的，光不稳定性的，或包括二硫键。
16.如权利要求9-15中任意一项的方法，其中，所述可检测的标记和/或可裂解的接头具有足以阻止第二个核苷酸或核苷掺入到所述核酸分子中的大小。
17.一种用于测定靶单链多核苷酸的序列的方法，包括监测互补核苷酸的依次掺入，其中，每个所述核苷酸具有通过可裂解的接头与可检测的标记连接的碱基，并且，其中，每一个掺入的核苷酸的身份是通过检测与所述碱基连接的标记确定的，并且随后除去所述标记。
18.一种用于确定靶单链多核苷酸的序列的方法，包括：
(a)提供核苷酸，其中，所述核苷酸具有通过可裂解的接头与可检测的标记连接的碱基，并且，其中，所述与每一种类型的核苷酸连接的可检测的标记，在检测时可以与用于标记其他类型的核苷酸的可检测标记区分开来；
(b)将核苷酸掺入到所述靶单链多核苷酸的互补体上；
(c)检测(b)的核苷酸的标记，以便确定掺入的核苷酸的类型；
(d)除去(b)的核苷酸的标记；和
(e)任选重复步骤(b)-(d)一次或多次；
以便测定靶单链多核苷酸的序列。
19.如权利要求17的方法，其中，让每一个核苷酸依次与所述靶接触，在添加下一个核苷酸之前除去未掺入的核苷酸，并且其中所述标记的检测和除去是在添加每一个核苷酸之后，或在添加所有四种核苷酸之后进行的。
20.如权利要求17的方法，其中，让所有核苷酸同时与所述靶接触，并且在检测之前除去未掺入的核苷酸，并且随后除去所述标记。
21.如权利要求17的方法，包括第一个步骤和第二个步骤，其中，在第一个步骤中，让包括四种核苷酸中的两种的第一种组合物与所述靶接触，并且在检测之前除去未掺入的核苷酸，并且随后除去所述标记，并且，其中，在第二个步骤中，让包括第一种组合物中所不包括的两种核苷酸的第二种组合物与所述靶接触，并且在检测之前除去未掺入的核苷酸，并且随后除去所述标记，并且，其中，任选重复所述第一个步骤和第二个步骤一次或多次。
22.如权利要求17的方法，包括第一个步骤和第二个步骤，其中，在第一个步骤中，让包括四种核苷酸中的一种的组合物与所述靶接触，并且在检测之前除去未掺入的核苷酸，并且随后除去所述标记，并且，其中，在第二个步骤中，让包括第一种组合物中所不包括的三种核苷酸的第二种组合物与所述靶接触，并且在检测之前除去未掺入的核苷酸，并且随后除去所述标记，并且，其中，任选重复所述第一个步骤和第二个步骤任选一次或多次。
23.如权利要求17的方法，包括第一个步骤和第二个步骤，其中，在第一个步骤中，将包括四种核苷酸中的三种的组合物与所述靶接触，并且在检测之前除去未掺入的核苷酸，并且随后除去所述标记，并且，其中，在第二个步骤中，让包括第一种组合物中所不包括的核苷酸的组合物与所述靶接触，并且在检测之前除去未掺入的核苷酸，并且随后除去所述标记，并且，其中，任选重复所述第一个步骤和第二个步骤一次或多次。
24.一种试剂盒，所述试剂盒包括：
(a)各个核苷酸，其中，每一个核苷酸具有通过可裂解的接头与可检测的标记连接的碱基，并且，其中，与每一种核苷酸连接的所述可检测的标记，在检测时可以与用于标记其他三种核苷酸的可检测标记区分开来；和
(b)它们的包装材料。
25.如权利要求21的试剂盒，所述试剂盒还包括酶和适合所述酶起作用的缓冲液。
CNA028278321A 2001-12-04 2002-12-04 标记过的核苷酸 Pending CN1617937A (zh)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
GBGB0129012.1A GB0129012D0 (en) 2001-12-04 2001-12-04 Labelled nucleotides
GB0129012.1 2001-12-04
US10/227,131 US7057026B2 (en) 2001-12-04 2002-08-23 Labelled nucleotides
US10/227,131 2002-08-23

Publications (1)

Publication Number Publication Date
CN1617937A true CN1617937A (zh) 2005-05-18

Family

ID=26246831

Family Applications (1)

Application Number Title Priority Date Filing Date
CNA028278321A Pending CN1617937A (zh) 2001-12-04 2002-12-04 标记过的核苷酸

Country Status (17)

Country Link
US (4) US7057026B2 (zh)
EP (4) EP3266791B1 (zh)
JP (1) JP2005511058A (zh)
KR (1) KR20050044668A (zh)
CN (1) CN1617937A (zh)
AU (1) AU2002349155A1 (zh)
BR (1) BR0214683A (zh)
CA (1) CA2469066A1 (zh)
CY (2) CY1118765T1 (zh)
DK (2) DK3266791T3 (zh)
ES (2) ES2620131T3 (zh)
HK (1) HK1248710A1 (zh)
IL (1) IL162170A0 (zh)
IS (1) IS7291A (zh)
PT (2) PT3266791T (zh)
SI (1) SI3266791T1 (zh)
WO (1) WO2003048387A2 (zh)

Cited By (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101631796A (zh) * 2007-03-09 2010-01-20 独立行政法人理化学研究所 具有由单核苷或单核苷酸衍生的结构的化合物、核酸、标记物以及核酸检测方法和试剂盒
CN103087131A (zh) * 2013-01-15 2013-05-08 上海交通大学 可逆终端及其合成和在dna合成测序中的用途
CN103484106A (zh) * 2013-09-05 2014-01-01 上海交通大学 四色荧光标记可逆终端及其在dna测序中的用途
US8871921B2 (en) 2008-09-03 2014-10-28 Quantumdx Group Ltd. Design, synthesis and use of synthetic nucleotides comprising charge mass tags
CN105377869A (zh) * 2013-03-15 2016-03-02 伊鲁米纳剑桥有限公司 修饰的核苷或核苷酸
US9410196B2 (en) 2008-09-03 2016-08-09 Quantumdx Group Limited Methods and kits for nucleic acid sequencing
CN106117288A (zh) * 2015-05-08 2016-11-16 桑特里莱恩科技控股公司 二硫键连接的可逆终止子
CN108822173A (zh) * 2018-06-26 2018-11-16 合肥华纳生物医药科技有限公司 一种荧光标记可剪切核苷酸、合成方法及其在dna测序中的用途
WO2019042016A1 (zh) * 2017-08-31 2019-03-07 深圳华大智造科技有限公司 一种核酸探针以及一种核酸测序方法
CN109562142A (zh) * 2016-04-04 2019-04-02 纽约哥伦比亚大学董事会 使用核苷酸类似物和拉曼检测的dna合成测序
US10759824B2 (en) 2008-09-03 2020-09-01 Quantumdx Group Limited Design, synthesis and use of synthetic nucleotides comprising charge mass tags
WO2021031109A1 (zh) * 2019-08-20 2021-02-25 深圳华大智造极创科技有限公司 一种基于发光标记物光信号动力学及二次发光信号对多核苷酸进行测序的方法
US11180523B2 (en) 2016-01-04 2021-11-23 Quantumdx Group Limited Design, synthesis and use of synthetic nucleotides comprising charge mass tags
CN114989235A (zh) * 2015-09-28 2022-09-02 哥伦比亚大学董事会 用作dna合成测序的可逆终止物的基于新的二硫键接头的核苷酸的设计与合成

Families Citing this family (748)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6780591B2 (en) 1998-05-01 2004-08-24 Arizona Board Of Regents Method of determining the nucleotide sequence of oligonucleotides and DNA molecules
US7875440B2 (en) 1998-05-01 2011-01-25 Arizona Board Of Regents Method of determining the nucleotide sequence of oligonucleotides and DNA molecules
ATE356222T1 (de) 2000-10-06 2007-03-15 Univ Columbia Massives parallelverfahren zur dekodierung von dna und rna
US9708358B2 (en) 2000-10-06 2017-07-18 The Trustees Of Columbia University In The City Of New York Massive parallel method for decoding DNA and RNA
GB0129012D0 (en) * 2001-12-04 2002-01-23 Solexa Ltd Labelled nucleotides
US7057026B2 (en) * 2001-12-04 2006-06-06 Solexa Limited Labelled nucleotides
US20030215816A1 (en) * 2002-05-20 2003-11-20 Narayan Sundararajan Method for sequencing nucleic acids by observing the uptake of nucleotides modified with bulky groups
HUE032483T2 (en) 2002-08-23 2017-09-28 Illumina Cambridge Ltd Marked nucleotides
US11008359B2 (en) 2002-08-23 2021-05-18 Illumina Cambridge Limited Labelled nucleotides
US7414116B2 (en) 2002-08-23 2008-08-19 Illumina Cambridge Limited Labelled nucleotides
SI3587433T1 (sl) 2002-08-23 2020-08-31 Illumina Cambridge Limited Modificirani nukleotidi
WO2004055160A2 (en) * 2002-12-13 2004-07-01 The Trustees Of Columbia University In The City Of New York Biomolecular coupling methods using 1,3-dipolar cycloaddition chemistry
US20060078893A1 (en) 2004-10-12 2006-04-13 Medical Research Council Compartmentalised combinatorial chemistry by microfluidic control
GB0307403D0 (en) 2003-03-31 2003-05-07 Medical Res Council Selection by compartmentalised screening
GB0307428D0 (en) 2003-03-31 2003-05-07 Medical Res Council Compartmentalised combinatorial chemistry
US7572581B2 (en) * 2003-06-30 2009-08-11 Roche Molecular Systems, Inc. 2′-terminator nucleotide-related methods and systems
EP1725572B1 (de) 2003-11-05 2017-05-31 AGCT GmbH Makromolekulare nukleotidverbindungen und methoden zu deren anwendung
US7169560B2 (en) 2003-11-12 2007-01-30 Helicos Biosciences Corporation Short cycle methods for sequencing polynucleotides
WO2005080605A2 (en) 2004-02-19 2005-09-01 Helicos Biosciences Corporation Methods and kits for analyzing polynucleotide sequences
WO2005084367A2 (en) * 2004-03-03 2005-09-15 The Trustees Of Columbia University In The City Of New York Photocleavable fluorescent nucleotides for dna sequencing on chip constructed by site-specific coupling chemistry
US20050221339A1 (en) 2004-03-31 2005-10-06 Medical Research Council Harvard University Compartmentalised screening by microfluidic control
WO2006073436A2 (en) * 2004-04-29 2006-07-13 The Trustees Of Columbia University In The City Of New York Mass tag pcr for multiplex diagnostics
US7476734B2 (en) * 2005-12-06 2009-01-13 Helicos Biosciences Corporation Nucleotide analogs
US7968287B2 (en) 2004-10-08 2011-06-28 Medical Research Council Harvard University In vitro evolution in microfluidic systems
GB0427236D0 (en) 2004-12-13 2005-01-12 Solexa Ltd Improved method of nucleotide detection
WO2006064199A1 (en) * 2004-12-13 2006-06-22 Solexa Limited Improved method of nucleotide detection
EP1879906A4 (en) * 2005-01-05 2009-07-22 Advanced Genetic Analysis Corp REVERSIBLE NUCLEOTIDE TERMINATORS AND USES THEREOF
EP2241637A1 (en) 2005-02-01 2010-10-20 AB Advanced Genetic Analysis Corporation Nucleic acid sequencing by performing successive cycles of duplex extension
EP2272983A1 (en) 2005-02-01 2011-01-12 AB Advanced Genetic Analysis Corporation Reagents, methods and libraries for bead-based sequencing
GB0507835D0 (en) * 2005-04-18 2005-05-25 Solexa Ltd Method and device for nucleic acid sequencing using a planar wave guide
US8623628B2 (en) 2005-05-10 2014-01-07 Illumina, Inc. Polymerases
GB0509508D0 (en) * 2005-05-10 2005-06-15 Solexa Ltd Improved polymerases
US20060281100A1 (en) * 2005-06-14 2006-12-14 Shen Gene G Thiotriphosphate nucleotide dye terminators
US9169510B2 (en) * 2005-06-21 2015-10-27 The Trustees Of Columbia University In The City Of New York Pyrosequencing methods and related compositions
GB0517097D0 (en) 2005-08-19 2005-09-28 Solexa Ltd Modified nucleosides and nucleotides and uses thereof
US7666593B2 (en) 2005-08-26 2010-02-23 Helicos Biosciences Corporation Single molecule sequencing of captured nucleic acids
WO2007053702A2 (en) 2005-10-31 2007-05-10 The Trustees Of Columbia University In The City Of New York Synthesis of four color 3'-o-allyl modified photocleavable fluorescent nucleotides and related methods
WO2007053719A2 (en) 2005-10-31 2007-05-10 The Trustees Of Columbia University In The City Of New York Chemically cleavable 3'-o-allyl-dntp-allyl-fluorophore fluorescent nucleotide analogues and related methods
CA2630544A1 (en) * 2005-11-21 2007-05-31 The Trustees Of Columbia University In The City Of New York Multiplex digital immuno-sensing using a library of photocleavable mass tags
WO2007081385A2 (en) 2006-01-11 2007-07-19 Raindance Technologies, Inc. Microfluidic devices and methods of use in the formation and control of nanoreactors
CN101400803B (zh) * 2006-01-19 2013-04-17 纽约州立大学研究基金会 用于检测和鉴定经编码的珠粒和生物分子的方法和装置
CA2648149A1 (en) * 2006-03-31 2007-11-01 Solexa, Inc. Systems and devices for sequence by synthesis analysis
WO2007114693A2 (en) 2006-04-04 2007-10-11 Keygene N.V. High throughput detection of molecular markers based on aflp and high throughput sequencing
JP2009538123A (ja) * 2006-04-19 2009-11-05 アプライド バイオシステムズ， エルエルシー ゲル非含有ビーズベースの配列決定のための試薬、方法およびライブラリー
EP2530168B1 (en) 2006-05-11 2015-09-16 Raindance Technologies, Inc. Microfluidic Devices
US9562837B2 (en) 2006-05-11 2017-02-07 Raindance Technologies, Inc. Systems for handling microfludic droplets
US9074242B2 (en) 2010-02-12 2015-07-07 Raindance Technologies, Inc. Digital analyte analysis
WO2007135368A2 (en) 2006-05-18 2007-11-29 Solexa Limited Dye compounds and the use of their labelled conjugates
WO2007146158A1 (en) * 2006-06-07 2007-12-21 The Trustees Of Columbia University In The City Of New York Dna sequencing by nanopore using modified nucleotides
WO2008005673A2 (en) * 2006-06-30 2008-01-10 Applera Corporation Reversible terminator nucleotides and methods of use
US8178300B2 (en) 2006-07-12 2012-05-15 Keygene N.V. Method for the identification of the clonal source of a restriction fragment
US9012390B2 (en) 2006-08-07 2015-04-21 Raindance Technologies, Inc. Fluorocarbon emulsion stabilizing surfactants
CA2660286A1 (en) 2006-08-09 2008-02-21 Homestead Clinical Corporation Organ-specific proteins and methods of their use
WO2008027558A2 (en) 2006-08-31 2008-03-06 Codon Devices, Inc. Iterative nucleic acid assembly using activation of vector-encoded traits
US8399188B2 (en) 2006-09-28 2013-03-19 Illumina, Inc. Compositions and methods for nucleotide sequencing
US8568979B2 (en) 2006-10-10 2013-10-29 Illumina, Inc. Compositions and methods for representational selection of nucleic acids from complex mixtures using hybridization
US20080242560A1 (en) * 2006-11-21 2008-10-02 Gunderson Kevin L Methods for generating amplified nucleic acid arrays
DE112007002932B4 (de) 2006-12-01 2015-08-06 The Trustees Of Columbia University In The City Of New York Vierfarben DNA-Sequenzierung mittels Synthese unter Verwendung von abspaltbaren, reversiblen, fluoreszierenden Nucleotidterminatoren
BRPI0719923B1 (pt) * 2006-12-05 2021-08-17 Agilent Technologies Inc Métodos para sequenciamento de um ácido nucléico alvo, para conversão de um componente de ocorrência não natural em uma molécula de ácido nucléico em um componente de ocorrência natural, para terminar uma síntese de ácido nucléico, e para realizar o sequenciamento de sanger ou tipo sanger, e compostos
US7897737B2 (en) 2006-12-05 2011-03-01 Lasergen, Inc. 3′-OH unblocked, nucleotides and nucleosides, base modified with photocleavable, terminating groups and methods for their use in DNA sequencing
EP2121983A2 (en) 2007-02-02 2009-11-25 Illumina Cambridge Limited Methods for indexing samples and sequencing multiple nucleotide templates
US11940413B2 (en) 2007-02-05 2024-03-26 IsoPlexis Corporation Methods and devices for sequencing nucleic acids in smaller batches
US11035823B2 (en) 2009-03-17 2021-06-15 Qiagen Sciences, Llc Methods and devices for sequencing nucleic acids in smaller batches
US8481259B2 (en) 2007-02-05 2013-07-09 Intelligent Bio-Systems, Inc. Methods and devices for sequencing nucleic acids in smaller batches
US8772046B2 (en) 2007-02-06 2014-07-08 Brandeis University Manipulation of fluids and reactions in microfluidic systems
JP5750209B2 (ja) * 2007-03-16 2015-07-15 アプタ バイオサイエンス リミテッド 機能性分子、機能性分子合成用アミダイド、及び標的物質解析方法
US8592221B2 (en) 2007-04-19 2013-11-26 Brandeis University Manipulation of fluids, fluid components and reactions in microfluidic systems
US7678894B2 (en) * 2007-05-18 2010-03-16 Helicos Biosciences Corporation Nucleotide analogs
US20090099027A1 (en) * 2007-08-23 2009-04-16 Applied Biosystems Inc. Methods of Modifying Support Surfaces for the Immobilization of Particles and the Use of the Immobilized Particles for Analyzing Nucleic Acids
US20090062132A1 (en) * 2007-08-29 2009-03-05 Borner Scott R Alternative nucleic acid sequencing methods
DK2725107T3 (da) 2007-10-19 2019-01-02 Univ Columbia DNA-sekventering med ikke-fluorescerende nukleotidreversible terminatorer og ddNTP'er modificeret med spaltbart mærke og nukleinsyre omfattende inosin med reversible terminatorer
EP2207900B1 (en) 2007-10-19 2015-04-29 The Trustees of Columbia University in the City of New York Design and synthesis of cleavable fluorescent nucleotides as reversible terminators for dna sequencing by synthesis
US7767441B2 (en) * 2007-10-25 2010-08-03 Industrial Technology Research Institute Bioassay system including optical detection apparatuses, and method for detecting biomolecules
US7811810B2 (en) 2007-10-25 2010-10-12 Industrial Technology Research Institute Bioassay system including optical detection apparatuses, and method for detecting biomolecules
US8017338B2 (en) * 2007-11-20 2011-09-13 Life Technologies Corporation Reversible di-nucleotide terminator sequencing
US8652781B2 (en) 2008-02-12 2014-02-18 Pacific Biosciences Of California, Inc. Cognate sampling kinetics
WO2011025477A1 (en) 2009-08-25 2011-03-03 Illumina, Inc. Methods for selecting and amplifying polynucleotides
US9695469B2 (en) 2008-03-17 2017-07-04 Stichting Genetwister Ip Expression-linked gene discovery
US9017973B2 (en) 2008-03-19 2015-04-28 Intelligent Biosystems, Inc. Methods and compositions for incorporating nucleotides
US8623598B2 (en) 2008-03-19 2014-01-07 Intelligent Bio Systems, Inc. Methods and compositions for inhibiting undesired cleaving of labels
US8420366B2 (en) * 2008-03-31 2013-04-16 Pacific Biosciences Of California, Inc. Generation of modified polymerases for improved accuracy in single molecule sequencing
EP2942404B1 (en) * 2008-03-31 2016-11-23 Pacific Biosciences of California, Inc. Generation of modified polymerases for improved accuracy in single molecule sequencing
US8133672B2 (en) * 2008-03-31 2012-03-13 Pacific Biosciences Of California, Inc. Two slow-step polymerase enzyme systems and methods
US8999676B2 (en) 2008-03-31 2015-04-07 Pacific Biosciences Of California, Inc. Recombinant polymerases for improved single molecule sequencing
US8039817B2 (en) 2008-05-05 2011-10-18 Illumina, Inc. Compensator for multiple surface imaging
US20110105356A1 (en) * 2008-05-07 2011-05-05 Derosier Chad F Compositions and methods for providing substances to and from an array
WO2009152353A2 (en) 2008-06-11 2009-12-17 Lasergen, Inc. Nucleotides and nucleosides and methods for their use in dna sequencing
WO2010003132A1 (en) 2008-07-02 2010-01-07 Illumina Cambridge Ltd. Using populations of beads for the fabrication of arrays on surfaces
US20100120034A1 (en) * 2008-07-03 2010-05-13 Life Technologies Corporation Methylation analysis of mate pairs
EP4047367A1 (en) 2008-07-18 2022-08-24 Bio-Rad Laboratories, Inc. Method for detecting target analytes with droplet libraries
US8173198B2 (en) * 2008-07-23 2012-05-08 Life Technologies Corporation Deposition of metal oxides onto surfaces as an immobilization vehicle for carboxylated or phophated particles or polymers
CA2735979A1 (en) * 2008-09-05 2010-03-11 Pacific Biosciences Of California, Inc. Engineering polymerases and reaction conditions for modified incorporation properties
US8383345B2 (en) 2008-09-12 2013-02-26 University Of Washington Sequence tag directed subassembly of short sequencing reads into long sequencing reads
WO2010038042A1 (en) 2008-10-02 2010-04-08 Illumina Cambridge Ltd. Nucleic acid sample enrichment for sequencing applications
EP2340314B8 (en) * 2008-10-22 2015-02-18 Illumina, Inc. Preservation of information related to genomic dna methylation
CA2742298C (en) * 2008-11-03 2019-09-10 The Regents Of The University Of California Methods for detecting modification resistant nucleic acids
US8252910B2 (en) 2008-11-19 2012-08-28 Pacific Biosciences Of California, Inc. Modular nucleotide compositions and uses therefor
WO2010068702A2 (en) * 2008-12-10 2010-06-17 Illumina, Inc. Methods and compositions for hybridizing nucleic acids
US8236532B2 (en) 2008-12-23 2012-08-07 Illumina, Inc. Multibase delivery for long reads in sequencing by synthesis protocols
CN102272334B (zh) 2009-01-13 2014-08-20 关键基因股份有限公司 新基因组测序策略
EP3415235A1 (en) 2009-03-23 2018-12-19 Raindance Technologies Inc. Manipulation of microfluidic droplets
WO2010111674A2 (en) 2009-03-27 2010-09-30 Life Technologies Corporation Methods and apparatus for single molecule sequencing using energy transfer detection
GB0907372D0 (en) 2009-04-29 2009-06-10 Invitrogen Dynal As Particles
CA2760439A1 (en) 2009-04-30 2010-11-04 Good Start Genetics, Inc. Methods and compositions for evaluating genetic markers
EP2427572B1 (en) * 2009-05-01 2013-08-28 Illumina, Inc. Sequencing methods
US8632975B2 (en) 2009-06-05 2014-01-21 Life Technologies Corporation Nucleotide transient binding for sequencing methods
DK2456892T3 (en) 2009-07-24 2014-12-08 Illumina Inc Procedure for sequencing of a polynukleotidskabelon
WO2011042564A1 (en) 2009-10-09 2011-04-14 Universite De Strasbourg Labelled silica-based nanomaterial with enhanced properties and uses thereof
US9315860B2 (en) 2009-10-26 2016-04-19 Genovoxx Gmbh Conjugates of nucleotides and method for the application thereof
EP2517025B1 (en) 2009-12-23 2019-11-27 Bio-Rad Laboratories, Inc. Methods for reducing the exchange of molecules between droplets
US8422031B2 (en) 2010-02-01 2013-04-16 Illumina, Inc. Focusing methods and optical systems and assemblies using the same
US20110192723A1 (en) * 2010-02-08 2011-08-11 Genia Technologies, Inc. Systems and methods for manipulating a molecule in a nanopore
US8324914B2 (en) 2010-02-08 2012-12-04 Genia Technologies, Inc. Systems and methods for characterizing a molecule
US9605307B2 (en) 2010-02-08 2017-03-28 Genia Technologies, Inc. Systems and methods for forming a nanopore in a lipid bilayer
US9678055B2 (en) 2010-02-08 2017-06-13 Genia Technologies, Inc. Methods for forming a nanopore in a lipid bilayer
US9366632B2 (en) 2010-02-12 2016-06-14 Raindance Technologies, Inc. Digital analyte analysis
US9399797B2 (en) 2010-02-12 2016-07-26 Raindance Technologies, Inc. Digital analyte analysis
US10351905B2 (en) 2010-02-12 2019-07-16 Bio-Rad Laboratories, Inc. Digital analyte analysis
US8759037B2 (en) 2010-02-23 2014-06-24 Illumina Cambridge Limited Amplification methods to minimise sequence specific bias
WO2011112465A1 (en) 2010-03-06 2011-09-15 Illumina, Inc. Systems, methods, and apparatuses for detecting optical signals from a sample
WO2011123246A2 (en) 2010-04-01 2011-10-06 Illumina, Inc. Solid-phase clonal amplification and related methods
CA2794522C (en) 2010-04-05 2019-11-26 Prognosys Biosciences, Inc. Spatially encoded biological assays
US20190300945A1 (en) 2010-04-05 2019-10-03 Prognosys Biosciences, Inc. Spatially Encoded Biological Assays
US10787701B2 (en) 2010-04-05 2020-09-29 Prognosys Biosciences, Inc. Spatially encoded biological assays
WO2011159942A1 (en) 2010-06-18 2011-12-22 Illumina, Inc. Conformational probes and methods for sequencing nucleic acids
US8986930B2 (en) 2010-07-12 2015-03-24 Pacific Biosciences Of California, Inc. Sequencing reactions with alkali metal cations for pulse width control
US9029103B2 (en) 2010-08-27 2015-05-12 Illumina Cambridge Limited Methods for sequencing polynucleotides
JP5917519B2 (ja) 2010-09-10 2016-05-18 バイオ−ラッド ラボラトリーズ インコーポレーティッド クロマチン分析のためのｄｎａのサイズ選択
US8483969B2 (en) 2010-09-17 2013-07-09 Illuminia, Inc. Variation analysis for multiple templates on a solid support
US8759038B2 (en) 2010-09-29 2014-06-24 Illumina Cambridge Limited Compositions and methods for sequencing nucleic acids
WO2012045012A2 (en) 2010-09-30 2012-04-05 Raindance Technologies, Inc. Sandwich assays in droplets
EP2633069B1 (en) 2010-10-26 2015-07-01 Illumina, Inc. Sequencing methods
EP3928867A1 (en) 2010-10-27 2021-12-29 Illumina, Inc. Microdevices and biosensor cartridges for biological or chemical analysis and systems and methods for the same
US8575071B2 (en) 2010-11-03 2013-11-05 Illumina, Inc. Reducing adapter dimer formation
CA2821299C (en) 2010-11-05 2019-02-12 Frank J. Steemers Linking sequence reads using paired code tags
US9074251B2 (en) 2011-02-10 2015-07-07 Illumina, Inc. Linking sequence reads using paired code tags
WO2012064975A1 (en) 2010-11-12 2012-05-18 Gen9, Inc. Protein arrays and methods of using and making the same
CA3132011A1 (en) 2010-11-12 2012-06-14 Gen9, Inc. Methods and devices for nucleic acids synthesis
WO2012074855A2 (en) 2010-11-22 2012-06-07 The Regents Of The University Of California Methods of identifying a cellular nascent rna transcript
WO2013082164A1 (en) 2011-11-28 2013-06-06 Life Technologies Corporation Enhanced ligation reactions
WO2012088341A2 (en) 2010-12-22 2012-06-28 Genia Technologies, Inc. Nanopore-based single dna molecule characterization, identification and isolation using speed bumps
US9163281B2 (en) 2010-12-23 2015-10-20 Good Start Genetics, Inc. Methods for maintaining the integrity and identification of a nucleic acid template in a multiplex sequencing reaction
US8951781B2 (en) 2011-01-10 2015-02-10 Illumina, Inc. Systems, methods, and apparatuses to image a sample for biological or chemical analysis
US8883764B1 (en) 2011-01-19 2014-11-11 Illumina, Inc. Formylpyrrole-based heterocycles for nucleic acid attachment to supports
US8962242B2 (en) 2011-01-24 2015-02-24 Genia Technologies, Inc. System for detecting electrical properties of a molecular complex
US9110478B2 (en) 2011-01-27 2015-08-18 Genia Technologies, Inc. Temperature regulation of measurement arrays
ES2627851T3 (es) 2011-01-31 2017-07-31 Illumina, Inc. Métodos para reducir el daño en ácidos nucleicos
WO2012106546A2 (en) 2011-02-02 2012-08-09 University Of Washington Through Its Center For Commercialization Massively parallel continguity mapping
EP3412778A1 (en) 2011-02-11 2018-12-12 Raindance Technologies, Inc. Methods for forming mixed droplets
WO2012112804A1 (en) 2011-02-18 2012-08-23 Raindance Technoligies, Inc. Compositions and methods for molecular labeling
GB201106254D0 (en) 2011-04-13 2011-05-25 Frisen Jonas Method and product
EP2702171A1 (de) 2011-04-27 2014-03-05 Cherkasov, Dmitry Methoden und komponenten zur detektion von nukleinsäureketten
US20150086981A1 (en) 2011-05-04 2015-03-26 Genovoxx Gmbh Nucleoside-triphosphate conjugate and methods for the use thereof
WO2012162429A2 (en) 2011-05-23 2012-11-29 The Trustees Of Columbia University In The City Of New York Dna sequencing by synthesis using raman and infrared spectroscopy detection
WO2012167142A2 (en) 2011-06-02 2012-12-06 Raindance Technolgies, Inc. Enzyme quantification
US8841071B2 (en) 2011-06-02 2014-09-23 Raindance Technologies, Inc. Sample multiplexing
WO2012170936A2 (en) 2011-06-09 2012-12-13 Illumina, Inc. Patterned flow-cells useful for nucleic acid analysis
EP2729580B1 (en) 2011-07-08 2015-09-16 Keygene N.V. Sequence based genotyping based on oligonucleotide ligation assays
US8658430B2 (en) 2011-07-20 2014-02-25 Raindance Technologies, Inc. Manipulating droplet size
JP2014526899A (ja) 2011-08-26 2014-10-09 ジェン９・インコーポレイテッド 核酸の高忠実度アセンブリのための組成物および方法
US8889860B2 (en) 2011-09-13 2014-11-18 Lasergen, Inc. 3′-OH unblocked, fast photocleavable terminating nucleotides and methods for nucleic acid sequencing
AU2012316218B2 (en) 2011-09-26 2016-03-17 Gen-Probe Incorporated Algorithms for sequence determinations
US10378051B2 (en) 2011-09-29 2019-08-13 Illumina Cambridge Limited Continuous extension and deblocking in reactions for nucleic acids synthesis and sequencing
CA2852665A1 (en) 2011-10-17 2013-04-25 Good Start Genetics, Inc. Analysis methods
US8778849B2 (en) 2011-10-28 2014-07-15 Illumina, Inc. Microarray fabrication system and method
WO2013070627A2 (en) 2011-11-07 2013-05-16 Illumina, Inc. Integrated sequencing apparatuses and methods of use
EP2788499B1 (en) 2011-12-09 2016-01-13 Illumina, Inc. Expanded radix for polymeric tags
US9279154B2 (en) 2011-12-21 2016-03-08 Illumina, Inc. Apparatus and methods for kinetic analysis and determination of nucleic acid sequences
WO2013117595A2 (en) 2012-02-07 2013-08-15 Illumina Cambridge Limited Targeted enrichment and amplification of nucleic acids on a support
EP2817418B1 (en) 2012-02-24 2017-10-11 Raindance Technologies, Inc. Labeling and sample preparation for sequencing
US8986629B2 (en) 2012-02-27 2015-03-24 Genia Technologies, Inc. Sensor circuit for controlling, detecting, and measuring a molecular complex
NO2694769T3 (zh) 2012-03-06 2018-03-03
US9150853B2 (en) 2012-03-21 2015-10-06 Gen9, Inc. Methods for screening proteins using DNA encoded chemical libraries as templates for enzyme catalysis
US20130261984A1 (en) 2012-03-30 2013-10-03 Illumina, Inc. Methods and systems for determining fetal chromosomal abnormalities
WO2013150902A1 (ja) * 2012-04-02 2013-10-10 独立行政法人理化学研究所 非天然型塩基を含むオリゴデオキシヌクレオチドの切断による5'末端リン酸化オリゴデオキシヌクレオチドの製造方法
MX337140B (es) 2012-04-03 2016-02-12 Illumina Inc Cabazal integrado de lectura optoelectrónica y cartucho fluído útil para secuenciación de ácidos nucleicos.
US8209130B1 (en) 2012-04-04 2012-06-26 Good Start Genetics, Inc. Sequence assembly
US8812422B2 (en) 2012-04-09 2014-08-19 Good Start Genetics, Inc. Variant database
US9444880B2 (en) 2012-04-11 2016-09-13 Illumina, Inc. Cloud computing environment for biological data
US20130274148A1 (en) 2012-04-11 2013-10-17 Illumina, Inc. Portable genetic detection and analysis system and method
US10227635B2 (en) 2012-04-16 2019-03-12 Molecular Loop Biosolutions, Llc Capture reactions
US10081807B2 (en) 2012-04-24 2018-09-25 Gen9, Inc. Methods for sorting nucleic acids and multiplexed preparative in vitro cloning
WO2013185137A1 (en) 2012-06-08 2013-12-12 Pacific Biosciences Of California, Inc. Modified base detection with nanopore sequencing
US9012022B2 (en) 2012-06-08 2015-04-21 Illumina, Inc. Polymer coatings
US8895249B2 (en) 2012-06-15 2014-11-25 Illumina, Inc. Kinetic exclusion amplification of nucleic acid libraries
CA2876095A1 (en) 2012-06-15 2013-12-19 Genia Technologies, Inc. Chip set-up and high-accuracy nucleic acid sequencing
WO2014004393A1 (en) * 2012-06-25 2014-01-03 Gen9, Inc. Methods for nucleic acid assembly and high throughput sequencing
US20150167084A1 (en) 2012-07-03 2015-06-18 Sloan Kettering Institute For Cancer Research Quantitative Assessment of Human T-Cell Repertoire Recovery After Allogeneic Hematopoietic Stem Cell Transplantation
ES2637538T3 (es) 2012-07-17 2017-10-13 Counsyl, Inc. Sistema y métodos para la detección de una variación genética
US9092401B2 (en) 2012-10-31 2015-07-28 Counsyl, Inc. System and methods for detecting genetic variation
US9422602B2 (en) 2012-08-15 2016-08-23 Bio-Rad Laboratories, Inc. Methods and compositions for determining nucleic acid degradation
NL2017959B1 (en) 2016-12-08 2018-06-19 Illumina Inc Cartridge assembly
EP3699577B1 (en) 2012-08-20 2023-11-08 Illumina, Inc. System for fluorescence lifetime based sequencing
US9150896B2 (en) 2012-09-06 2015-10-06 Illumina, Inc. Nucleotides and primers with removable blocking groups
JP2015537190A (ja) 2012-09-19 2015-12-24 アッヴィ・バイオセラピューティクス・インコーポレイテッド 低減された免疫原性を有する抗体を同定するための方法
US9399766B2 (en) 2012-10-01 2016-07-26 Pacific Biosciences Of California, Inc. Recombinant polymerases for incorporation of protein shield nucleotide analogs
US9181583B2 (en) 2012-10-23 2015-11-10 Illumina, Inc. HLA typing using selective amplification and sequencing
US9116139B2 (en) 2012-11-05 2015-08-25 Illumina, Inc. Sequence scheduling and sample distribution techniques
US9605309B2 (en) 2012-11-09 2017-03-28 Genia Technologies, Inc. Nucleic acid sequencing using tags
EP2920725B1 (en) 2012-11-19 2021-11-10 Apton Biosystems, Inc. Digital analysis of molecular analytes using single molecule detection
US10829816B2 (en) 2012-11-19 2020-11-10 Apton Biosystems, Inc. Methods of analyte detection
US9683230B2 (en) 2013-01-09 2017-06-20 Illumina Cambridge Limited Sample preparation on a solid support
US9805407B2 (en) 2013-01-25 2017-10-31 Illumina, Inc. Methods and systems for using a cloud computing environment to configure and sell a biological sample preparation cartridge and share related data
US9759711B2 (en) 2013-02-05 2017-09-12 Genia Technologies, Inc. Nanopore arrays
US9512422B2 (en) 2013-02-26 2016-12-06 Illumina, Inc. Gel patterned surfaces
US9650406B2 (en) 2013-02-28 2017-05-16 Centrillion Technology Holdings Corporation Reversible terminator molecules and methods of their use
US9914979B2 (en) 2013-03-04 2018-03-13 Fry Laboratories, LLC Method and kit for characterizing microorganisms
US9873913B2 (en) 2013-03-08 2018-01-23 Roche Molecular Systems, Inc. Mutation testing
PL2969479T3 (pl) 2013-03-13 2021-12-27 Illumina, Inc. Wielowarstwowe urządzenia płynowe i sposoby ich wytwarzania
ES2887177T3 (es) 2013-03-13 2021-12-22 Illumina Inc Método de preparación de una biblioteca de secuenciación de ácidos nucleicos
US8778609B1 (en) 2013-03-14 2014-07-15 Good Start Genetics, Inc. Methods for analyzing nucleic acids
US10421996B2 (en) 2013-03-14 2019-09-24 Illumina, Inc. Modified polymerases for improved incorporation of nucleotide analogues
US9146248B2 (en) 2013-03-14 2015-09-29 Intelligent Bio-Systems, Inc. Apparatus and methods for purging flow cells in nucleic acid sequencing instruments
WO2014142981A1 (en) 2013-03-15 2014-09-18 Illumina, Inc. Enzyme-linked nucleotides
WO2014144883A1 (en) 2013-03-15 2014-09-18 The Trustees Of Columbia University In The City Of New York Raman cluster tagged molecules for biological imaging
US9255265B2 (en) 2013-03-15 2016-02-09 Illumina, Inc. Methods for producing stranded cDNA libraries
US20140274747A1 (en) 2013-03-15 2014-09-18 Illumina, Inc. Super resolution imaging
US9591268B2 (en) 2013-03-15 2017-03-07 Qiagen Waltham, Inc. Flow cell alignment methods and systems
US9193998B2 (en) 2013-03-15 2015-11-24 Illumina, Inc. Super resolution imaging
EP3005200A2 (en) 2013-06-03 2016-04-13 Good Start Genetics, Inc. Methods and systems for storing sequence read data
US9879313B2 (en) 2013-06-25 2018-01-30 Prognosys Biosciences, Inc. Methods and systems for determining spatial patterns of biological targets in a sample
KR102266002B1 (ko) 2013-07-01 2021-06-16 일루미나, 인코포레이티드 촉매-무함유 표면 작용화 및 중합체 그라프팅
ES2719579T3 (es) 2013-07-03 2019-07-11 Illumina Inc Sistema para secuenciación por síntesis ortogonal
US9410977B2 (en) 2013-08-08 2016-08-09 Illumina, Inc. Fluidic system for reagent delivery to a flow cell
EP4306650A3 (en) 2013-08-22 2024-03-27 Pacific Biosciences of California, Inc. Digital analysis of molecular analytes using electrical methods
CA2920390A1 (en) 2013-08-30 2015-03-05 Illumina, Inc. Manipulation of droplets on hydrophilic or variegated-hydrophilic surfaces
US9352315B2 (en) 2013-09-27 2016-05-31 Taiwan Semiconductor Manufacturing Company, Ltd. Method to produce chemical pattern in micro-fluidic structure
US11901041B2 (en) 2013-10-04 2024-02-13 Bio-Rad Laboratories, Inc. Digital analysis of nucleic acid modification
CN105683980B (zh) 2013-10-04 2018-08-24 生命科技股份有限公司 在使用终止化学物质的测序中建立分阶段效应模型的方法和系统
US9551697B2 (en) 2013-10-17 2017-01-24 Genia Technologies, Inc. Non-faradaic, capacitively coupled measurement in a nanopore cell array
US11041203B2 (en) 2013-10-18 2021-06-22 Molecular Loop Biosolutions, Inc. Methods for assessing a genomic region of a subject
US10851414B2 (en) 2013-10-18 2020-12-01 Good Start Genetics, Inc. Methods for determining carrier status
CA2926138A1 (en) 2013-10-23 2015-04-30 Genia Technologies, Inc. High speed molecular sensing with nanopores
US9322062B2 (en) 2013-10-23 2016-04-26 Genia Technologies, Inc. Process for biosensor well formation
US10540783B2 (en) 2013-11-01 2020-01-21 Illumina, Inc. Image analysis useful for patterned objects
CN110452809B (zh) 2013-11-17 2022-07-01 宽腾矽公司 与测定芯片通过接口连接的仪器
EP2876166B1 (en) 2013-11-20 2016-12-14 Roche Diagnostics GmbH New compound for sequencing by synthesis
WO2015084985A2 (en) 2013-12-03 2015-06-11 Illumina, Inc. Methods and systems for analyzing image data
DK3080585T3 (da) 2013-12-10 2024-02-05 Illumina Inc Biosensorer til biologisk eller kemisk analyse og fremgangsmåder til fremstilling heraf
US9944977B2 (en) 2013-12-12 2018-04-17 Raindance Technologies, Inc. Distinguishing rare variations in a nucleic acid sequence from a sample
DK3083994T3 (da) 2013-12-20 2021-09-13 Illumina Inc Bevarelse af genomisk konnektivitetsinformation i fragmenterede genomiske DNA-prøver
US20180073065A1 (en) 2013-12-23 2018-03-15 Illumina, Inc. Structured substrates for improving detection of light emissions and methods relating to the same
US10537889B2 (en) 2013-12-31 2020-01-21 Illumina, Inc. Addressable flow cell using patterned electrodes
EP3090063B1 (en) 2013-12-31 2019-11-06 Bio-Rad Laboratories, Inc. Method for detection of latent retrovirus
KR102001554B1 (ko) 2014-01-16 2019-07-18 일루미나, 인코포레이티드 고형 지지체 상에서의 앰플리콘 제조 방법 및 시퀀싱
US9677132B2 (en) 2014-01-16 2017-06-13 Illumina, Inc. Polynucleotide modification on solid support
CN110955371B (zh) 2014-02-13 2023-09-12 Illumina公司 综合式消费者基因组服务
CA2940048C (en) 2014-02-18 2023-03-14 Illumina, Inc. Methods and compositions for dna profiling
EP3116651B1 (en) 2014-03-11 2020-04-22 Illumina, Inc. Disposable, integrated microfluidic cartridge and methods of making it
FR3020071B1 (fr) 2014-04-17 2017-12-22 Dna Script Procede de synthese d'acides nucleiques, notamment d'acides nucleiques de grande longueur, utilisation du procede et kit pour la mise en œuvre du procede
WO2015168161A2 (en) 2014-04-29 2015-11-05 Illumina, Inc. Multiplexed single cell gene expression analysis using template switch and tagmentation
US11053548B2 (en) 2014-05-12 2021-07-06 Good Start Genetics, Inc. Methods for detecting aneuploidy
CA3219413A1 (en) 2014-05-16 2015-11-19 Illumina, Inc. Nucleic acid synthesis techniques
BR112016027815B1 (pt) 2014-05-27 2022-07-12 Illumina, Inc. Sistemas e métodos para análise bioquímica incluindo um instrumento de base e cartucho removível
WO2015187670A2 (en) 2014-06-03 2015-12-10 Illumina, Inc. Compositions, systems, and methods for detecting events using tethers anchored to or adjacent to nanopores
US20150353989A1 (en) 2014-06-09 2015-12-10 Illumina Cambridge Limited Sample preparation for nucleic acid amplification
WO2015189636A1 (en) 2014-06-13 2015-12-17 Illumina Cambridge Limited Methods and compositions for preparing sequencing libraries
US10017759B2 (en) 2014-06-26 2018-07-10 Illumina, Inc. Library preparation of tagged nucleic acid
CA2951416A1 (en) 2014-06-27 2015-12-30 Illumina, Inc. Modified polymerases for improved incorporation of nucleotide analogues
CA2953791A1 (en) 2014-06-30 2016-01-07 Illumina, Inc. Methods and compositions using one-sided transposition
PL3169805T3 (pl) 2014-07-15 2019-01-31 Illumina, Inc. Aktywowane biochemicznie urządzenie elektroniczne
US10457969B2 (en) 2014-07-21 2019-10-29 Illumina, Inc. Polynucleotide enrichment using CRISPR-Cas systems
WO2016023011A1 (en) 2014-08-08 2016-02-11 Quantum-Si Incorporated Integrated device with external light source for probing, detecting, and analyzing molecules
GB201414098D0 (en) 2014-08-08 2014-09-24 Illumina Cambridge Ltd Modified nucleotide linkers
AU2015300766B2 (en) 2014-08-08 2021-02-04 Quantum-Si Incorporated Integrated device for temporal binning of received photons
BR112017002489B1 (pt) 2014-08-08 2023-02-14 Quantum-Si Incorporated Instrumento configurado para fazer interface com um chip de teste, aparelho, método de análise de um espécime, método para sequenciar uma molécula-alvo de ácido nucleico e método para sequenciamento de ácido nucleico
WO2016026924A1 (en) 2014-08-21 2016-02-25 Illumina Cambridge Limited Reversible surface functionalization
FR3025201B1 (fr) 2014-09-02 2018-10-12 Dna Script Nucleotides modifies pour la synthese d'acides nucleiques, un kit renfermant de tels nucleotides et leur utilisation pour la production de genes ou sequences d'acides nucleiques synthetiques
WO2016040446A1 (en) 2014-09-10 2016-03-17 Good Start Genetics, Inc. Methods for selectively suppressing non-target sequences
WO2016040602A1 (en) 2014-09-11 2016-03-17 Epicentre Technologies Corporation Reduced representation bisulfite sequencing using uracil n-glycosylase (ung) and endonuclease iv
EP3191606B1 (en) 2014-09-12 2020-05-27 Illumina, Inc. Methods for detecting the presence of polymer subunits using chemiluminescence
US20160085910A1 (en) 2014-09-18 2016-03-24 Illumina, Inc. Methods and systems for analyzing nucleic acid sequencing data
EP3224595A4 (en) 2014-09-24 2018-06-13 Good Start Genetics, Inc. Process control for increased robustness of genetic assays
EP3201355B1 (en) 2014-09-30 2019-07-31 Illumina, Inc. Modified polymerases for improved incorporation of nucleotide analogues
US10487357B2 (en) 2014-10-03 2019-11-26 Life Technologies Corporation Methods of nucleic acid analysis using terminator nucleotides
US10544455B2 (en) 2014-10-03 2020-01-28 Life Technologies Corporation Sequencing methods, compositions and systems using terminator nucleotides
AU2015330688B2 (en) 2014-10-09 2021-02-11 Illumina, Inc. Method and device for separating immiscible liquids to effectively isolate at least one of the liquids
US9897791B2 (en) 2014-10-16 2018-02-20 Illumina, Inc. Optical scanning systems for in situ genetic analysis
WO2016061517A2 (en) 2014-10-17 2016-04-21 Illumina Cambridge Limited Contiguity preserving transposition
EP3699283A1 (en) 2014-10-20 2020-08-26 Molecular Assemblies Inc. Modified template-independent enzymes for polydeoxynucleotide systhesis
US10190162B2 (en) 2014-10-23 2019-01-29 Complete Genomics, Inc. Signal confinement sequencing (SCS) and nucleotide analogues for signal confinement sequencing
CA2965578C (en) 2014-10-31 2024-03-19 Illumina Cambridge Limited Polymers and dna copolymer coatings
US10000799B2 (en) 2014-11-04 2018-06-19 Boreal Genomics, Inc. Methods of sequencing with linked fragments
GB201419731D0 (en) 2014-11-05 2014-12-17 Illumina Cambridge Ltd Sequencing from multiple primers to increase data rate and density
CN107250358B (zh) 2014-11-05 2021-03-30 伊卢米纳剑桥有限公司 在样品制备和测序期间使用铁载体螯合剂减少dna损伤
MY182476A (en) 2014-11-11 2021-01-25 Illumina Cambridge Ltd Methods and arrays for producing and sequencing monoclonal clusters of nucleic acid
US10577649B2 (en) 2014-11-11 2020-03-03 Illumina, Inc. Polynucleotide amplification using CRISPR-Cas systems
EP3227684B1 (en) 2014-12-03 2019-10-02 Isoplexis Corporation Analysis and screening of cell secretion profiles
PT3234187T (pt) 2014-12-15 2021-05-26 Illumina Inc Método para colocação de moleculas de alvo unico num substrato
EP3271480B8 (en) 2015-01-06 2022-09-28 Molecular Loop Biosciences, Inc. Screening for structural variants
KR20210135626A (ko) 2015-02-10 2021-11-15 일루미나, 인코포레이티드 세포 성분을 분석하기 위한 방법 및 조성물
CN107847930B (zh) 2015-03-20 2020-06-30 亿明达股份有限公司 在竖直或大致竖直的位置中使用的流体盒
US10300452B2 (en) 2015-03-24 2019-05-28 Pacific Biosciences Of California, Inc. Methods and compositions for single molecule composition loading
WO2016154193A1 (en) 2015-03-24 2016-09-29 Illumina, Inc. Methods, carrier assemblies, and systems for imaging samples for biological or chemical analysis
ES2846730T3 (es) 2015-03-31 2021-07-29 Illumina Cambridge Ltd Concatemerización en superficie de moldes
EP4321627A3 (en) 2015-04-10 2024-04-17 10x Genomics Sweden AB Spatially distinguished, multiplex nucleic acid analysis of biological specimens
US10900030B2 (en) 2015-04-14 2021-01-26 Illumina, Inc. Structured substrates for improving detection of light emissions and methods relating to the same
US10844428B2 (en) 2015-04-28 2020-11-24 Illumina, Inc. Error suppression in sequenced DNA fragments using redundant reads with unique molecular indices (UMIS)
EP3760737B1 (en) 2015-05-11 2023-02-15 Illumina, Inc. Platform for discovery and analysis of therapeutic agents
CA2984112C (en) 2015-05-12 2021-07-06 Illumina, Inc. Field-effect apparatus and methods for sequencing nucelic acids
US10174363B2 (en) 2015-05-20 2019-01-08 Quantum-Si Incorporated Methods for nucleic acid sequencing
WO2016196358A1 (en) 2015-05-29 2016-12-08 Epicentre Technologies Corporation Methods of analyzing nucleic acids
RU2699612C2 (ru) 2015-05-29 2019-09-06 Иллюмина, Инк. Кассета для образцов и аналитическая система для проведения определенных реакций
CN107849598B (zh) 2015-05-29 2021-12-14 伊卢米纳剑桥有限公司 簇中的表面引物的增强利用
AU2016270887B2 (en) 2015-06-03 2019-09-19 Illumina, Inc. Compositions, systems, and methods for sequencing polynucleotides using tethers anchored to polymerases adjacent to nanopores
WO2016201142A1 (en) 2015-06-09 2016-12-15 Life Technologies Corporation Methods, systems, compositions, kits, apparatus and computer-readable media for molecular tagging
EP3310933B1 (en) 2015-06-17 2020-11-18 The Translational Genomics Research Institute Methods for obtaining biological molecules from a sample
WO2017007757A1 (en) 2015-07-06 2017-01-12 Illumina, Inc. Balanced ac modulation for driving droplet operations electrodes
WO2017006108A1 (en) 2015-07-06 2017-01-12 Illumina Cambridge Limited Sample preparation for nucleic acid amplification
CN107924121B (zh) 2015-07-07 2021-06-08 亿明达股份有限公司 经由纳米压印的选择性表面图案化
ES2945607T3 (es) 2015-07-17 2023-07-04 Illumina Inc Láminas de polímero para aplicaciones de secuenciación
CA2993002A1 (en) 2015-07-20 2017-01-26 Sentilus Holdco, Llc Chips, detectors, and methods of making and using the same
US10913975B2 (en) 2015-07-27 2021-02-09 Illumina, Inc. Spatial mapping of nucleic acid sequence information
BR112017023418A2 (pt) 2015-07-30 2018-07-24 Illumina, Inc. desbloqueio ortogonal de nucleotídeos
KR20180040669A (ko) 2015-08-14 2018-04-20 일루미나, 인코포레이티드 유전 특징을 결정하기 위한 자기-반응성 센서를 사용하는 시스템 및 방법
WO2017034868A1 (en) 2015-08-24 2017-03-02 Illumina, Inc. In-line pressure accumulator and flow-control system for biological or chemical assays
CA3172078A1 (en) 2015-08-28 2017-03-09 Illumina, Inc. Nucleic acid sequence analysis from single cells
CN107921432A (zh) 2015-09-02 2018-04-17 伊卢米纳剑桥有限公司 改善流控系统中的液滴操作的系统和方法
US10647981B1 (en) 2015-09-08 2020-05-12 Bio-Rad Laboratories, Inc. Nucleic acid library generation methods and compositions
US10450598B2 (en) 2015-09-11 2019-10-22 Illumina, Inc. Systems and methods for obtaining a droplet having a designated concentration of a substance-of-interest
JP6657721B2 (ja) 2015-09-30 2020-03-04 ソニー株式会社 細胞の分析方法、細胞分析用チップ、細胞分析用試薬、細胞分析用キット及び細胞分析用装置
US10577643B2 (en) 2015-10-07 2020-03-03 Illumina, Inc. Off-target capture reduction in sequencing techniques
US10465232B1 (en) 2015-10-08 2019-11-05 Trace Genomics, Inc. Methods for quantifying efficiency of nucleic acid extraction and detection
RU2719991C2 (ru) 2015-10-22 2020-04-23 Иллюмина, Инк. Наполняющая текучая среда для струйных устройств
US11421264B2 (en) 2015-11-06 2022-08-23 IsoPlexis Corporation Thiol-containing cleave reagents and oxidative wash
KR102454782B1 (ko) * 2015-11-06 2022-10-13 퀴아젠 사이언시스, 엘엘씨 뉴클레오티드 유사체
WO2017083437A1 (en) 2015-11-09 2017-05-18 Radiaction Ltd. Radiation shielding apparatuses and applications thereof
WO2017087724A1 (en) 2015-11-17 2017-05-26 Omniome, Inc. Methods for determining sequence profiles
US10731211B2 (en) 2015-11-18 2020-08-04 Pacific Biosciences Of California, Inc. Methods and compositions for loading of polymerase complexes
CN116240272A (zh) 2015-11-19 2023-06-09 赛纳生物科技(北京)有限公司 一种用于获得多核苷酸的序列信息的试剂盒或系统
DK3384046T3 (da) 2015-12-01 2021-07-12 Illumina Inc Digitalt mikrofluidisk system til enkeltcelleisolering og karakterisering af analytter
CN109072300B (zh) 2015-12-17 2023-01-31 伊路敏纳公司 区分复杂生物样品中的甲基化水平
WO2017123533A1 (en) 2016-01-11 2017-07-20 Illumina, Inc. Detection apparatus having a microfluorometer, a fluidic system, and a flow cell latch clamp module
KR20180111999A (ko) 2016-02-17 2018-10-11 테서렉트 헬스, 인코포레이티드 수명 촬상 및 검출 응용을 위한 센서 및 디바이스
EP4053545A1 (en) 2016-03-24 2022-09-07 Illumina, Inc. Photonic superlattice-based devices and compositions for use in luminescent imaging, and methods of using the same
JP7086855B2 (ja) 2016-03-28 2022-06-20 エヌキャン ジェノミクス， インコーポレイテッド 連結された二重鎖標的捕捉
DK3377226T3 (da) 2016-03-28 2021-04-26 Illumina Inc Mikrorækker i flere niveauer
US10961573B2 (en) 2016-03-28 2021-03-30 Boreal Genomics, Inc. Linked duplex target capture
US11326206B2 (en) 2016-04-07 2022-05-10 Pacific Biosciences Of California, Inc. Methods of quantifying target nucleic acids and identifying sequence variants
WO2017176896A1 (en) 2016-04-07 2017-10-12 Illumina, Inc. Methods and systems for construction of normalized nucleic acid libraries
CN113916788A (zh) 2016-04-22 2022-01-11 伊鲁米那股份有限公司 在发光成像中使用的设备和组成物及使用其的方法
CA3011635C (en) 2016-04-22 2021-10-26 Omniome, Inc. Nucleic acid sequencing method and system employing enhanced detection of nucleotide-specific ternary complex formation
WO2017190018A1 (en) 2016-04-29 2017-11-02 Omniome, Inc. Sequencing method employing ternary complex destabilization to identify cognate nucleotides
CN109154024B (zh) 2016-04-29 2022-03-04 欧姆尼欧美公司 核酸序列测定方法
US10597643B2 (en) 2016-04-29 2020-03-24 Omniome, Inc. Polymerases engineered to reduce nucleotide-independent DNA binding
US11542544B2 (en) 2016-05-11 2023-01-03 Illumina, Inc. Polynucleotide enrichment and amplification using CRISPR-Cas or Argonaute systems
KR102490693B1 (ko) * 2016-05-16 2023-01-19 나노스트링 테크놀로지스, 인크. 샘플 중 표적 핵산을 검출하는 방법
WO2017201198A1 (en) 2016-05-18 2017-11-23 Illumina, Inc. Self assembled patterning using patterned hydrophobic surfaces
EP3463381A4 (en) * 2016-05-23 2020-01-01 The Trustees of Columbia University in the City of New York NUCLEOTIDE DERIVATIVES AND THEIR METHODS OF USE
US11299769B2 (en) 2016-06-06 2022-04-12 Redvault Biosciences, Lp Target reporter constructs and uses thereof
WO2018018008A1 (en) 2016-07-22 2018-01-25 Oregon Health & Science University Single cell whole genome libraries and combinatorial indexing methods of making thereof
WO2018034780A1 (en) 2016-08-15 2018-02-22 Omniome, Inc. Sequencing method for rapid identification and processing of cognate nucleotide pairs
EP4039824A1 (en) 2016-08-15 2022-08-10 Pacific Biosciences of California, Inc. Method and system for sequencing nucleic acids
WO2018057770A1 (en) 2016-09-22 2018-03-29 Illumina, Inc. Somatic copy number variation detection
WO2018064116A1 (en) 2016-09-28 2018-04-05 Illumina, Inc. Methods and systems for data compression
TWI781669B (zh) 2016-10-14 2022-10-21 美商伊路米納有限公司 卡匣總成
PT3529380T (pt) 2016-10-19 2022-08-31 Illumina Inc Métodos para ligação química de ácidos nucléicos
JP7091325B2 (ja) * 2016-11-04 2022-06-27 キアゲン サイエンシーズ， エルエルシー チオール含有切断試薬および酸化的洗浄液
DK3538891T3 (da) 2016-11-11 2022-03-28 Isoplexis Corp Sammensætninger og fremgangsmåder til samtidig genomisk, transkriptomisk og proteomisk analyse af enkeltceller
SG10201912285UA (en) 2016-11-16 2020-02-27 Illumina Inc Validation methods and systems for sequence variant calls
GB201619458D0 (en) 2016-11-17 2017-01-04 Spatial Transcriptomics Ab Method for spatial tagging and analysing nucleic acids in a biological specimen
CA3043489A1 (en) 2016-11-21 2018-05-24 Nanostring Technologies, Inc. Chemical compositions and methods of using same
EP3545284A4 (en) 2016-11-22 2020-07-01 Isoplexis Corporation SYSTEMS, DEVICES AND METHODS FOR CAPTURING CELLS AND MANUFACTURING METHODS THEREOF
EP4119663A1 (en) 2016-12-09 2023-01-18 The Broad Institute, Inc. Crispr effector system based diagnostics
US11268137B2 (en) 2016-12-09 2022-03-08 Boreal Genomics, Inc. Linked ligation
CA3046532A1 (en) 2016-12-22 2018-06-28 Illumina, Inc. Arrays including a resin film and a patterned polymer layer
MX2019006616A (es) 2016-12-22 2019-10-24 Illumina Inc Arreglo que incluye cebador secuenciador y entidad no secuenciadora.
WO2018119347A1 (en) 2016-12-22 2018-06-28 Quantum-Si Incorporated Integrated photodetector with direct binning pixel
JP7126501B2 (ja) 2016-12-22 2022-08-26 イラミーナ インコーポレーテッド 品質管理トレーサー付きアレイ
WO2018125759A1 (en) 2016-12-30 2018-07-05 Omniome, Inc. Method and system employing distinguishable polymerases for detecting ternary complexes and identifying cognate nucleotides
GB201704754D0 (en) 2017-01-05 2017-05-10 Illumina Inc Kinetic exclusion amplification of nucleic acid libraries
CA3049142A1 (en) 2017-01-06 2018-07-12 Illumina, Inc. Phasing correction
WO2018128544A1 (en) 2017-01-06 2018-07-12 Agendia N.V. Biomarkers for selecting patient groups, and uses thereof.
AU2018208462B2 (en) 2017-01-10 2021-07-29 Pacific Biosciences Of California, Inc. Polymerases engineered to reduce nucleotide-independent DNA binding
CA3045498C (en) 2017-01-17 2021-07-13 Illumina, Inc. Oncogenic splice variant determination
BR112019014651A2 (pt) 2017-01-18 2020-07-21 Illumina, Inc. métodos para sequenciar moléculas de ácido nucleico e para preparar adaptadores de sequenciamento, produto de programa de computador, e, sistema de computador.
WO2018136117A1 (en) 2017-01-20 2018-07-26 Omniome, Inc. Allele-specific capture of nucleic acids
GB201700983D0 (en) 2017-01-20 2017-03-08 Life Tech As Polymeric particles
AU2017394645B2 (en) 2017-01-20 2020-01-23 Pacific Biosciences Of California, Inc. Genotyping by polymerase binding
CA3050695C (en) 2017-01-20 2024-02-20 Omniome, Inc. Process for cognate nucleotide detection in a nucleic acid sequencing workflow
GB201701688D0 (en) 2017-02-01 2017-03-15 Illumia Inc System and method with fiducials in non-recliner layouts
GB201701686D0 (en) 2017-02-01 2017-03-15 Illunina Inc System & method with fiducials having offset layouts
GB201701689D0 (en) 2017-02-01 2017-03-15 Illumia Inc System and method with fiducials of non-closed shapes
US11492666B2 (en) 2017-02-15 2022-11-08 Pacific Biosciences Of California, Inc. Distinguishing sequences by detecting polymerase dissociation
CN109642259A (zh) 2017-02-17 2019-04-16 阿姆斯特丹自由大学医学中心基金会 使用肿瘤教育的血小板的针对癌症的群智能增强的诊断和治疗选择
BR112018076259A2 (pt) 2017-02-21 2019-03-26 Illumina, Inc. tagmentação usando transposomas imobilizados com ligantes
WO2018165207A1 (en) 2017-03-06 2018-09-13 Singular Genomic Systems, Inc. Nucleic acid sequencing-by-synthesis (sbs) methods that combine sbs cycle steps
US11104937B2 (en) 2017-03-15 2021-08-31 The Broad Institute, Inc. CRISPR effector system based diagnostics
US11021740B2 (en) 2017-03-15 2021-06-01 The Broad Institute, Inc. Devices for CRISPR effector system based diagnostics
KR20190140918A (ko) 2017-03-15 2019-12-20 더 브로드 인스티튜트, 인코퍼레이티드 바이러스 검출을 위한 crispr 이펙터 시스템 기반 진단
US11174515B2 (en) 2017-03-15 2021-11-16 The Broad Institute, Inc. CRISPR effector system based diagnostics
CA3056765C (en) 2017-03-17 2024-04-02 Apton Biosystems, Inc. Sequencing and high resolution imaging
CN110612351B (zh) 2017-03-20 2023-08-11 Illumina公司 用于制备核酸文库的方法和组合物
EP4053294A1 (en) 2017-03-24 2022-09-07 Life Technologies Corporation Polynucleotide adapters and methods of use thereof
US20210285041A1 (en) * 2017-03-28 2021-09-16 The Trustees Of Columbia University In The City Of New York 3'-o-modified nucleotide analogues with different cleavable linkers for attaching fluorescent labels to the base for dna sequencing by synthesis
EP3607087A4 (en) 2017-04-04 2020-12-30 Omniome, Inc. FLUIDIC APPARATUS AND USEFUL METHODS FOR CHEMICAL AND BIOLOGICAL REACTIONS
US10934584B2 (en) 2017-04-23 2021-03-02 Illumina, Inc. Compositions and methods for improving sample identification in indexed nucleic acid libraries
CA3059840C (en) 2017-04-23 2022-04-26 Illumina Cambridge Limited Compositions and methods for improving sample identification in indexed nucleic acid libraries
AU2018259202B2 (en) 2017-04-23 2022-03-24 Illumina Cambridge Limited Compositions and methods for improving sample identification in indexed nucleic acid libraries
US10995369B2 (en) 2017-04-23 2021-05-04 Illumina, Inc. Compositions and methods for improving sample identification in indexed nucleic acid libraries
US9951385B1 (en) 2017-04-25 2018-04-24 Omniome, Inc. Methods and apparatus that increase sequencing-by-binding efficiency
US10161003B2 (en) 2017-04-25 2018-12-25 Omniome, Inc. Methods and apparatus that increase sequencing-by-binding efficiency
CA3220983A1 (en) 2017-05-01 2018-11-08 Illumina, Inc. Optimal index sequences for multiplex massively parallel sequencing
US11028436B2 (en) 2017-05-08 2021-06-08 Illumina, Inc. Universal short adapters for indexing of polynucleotide samples
EP4293122A3 (en) 2017-06-07 2024-01-24 Oregon Health & Science University Single cell whole genome libraries for methylation sequencing
US11542540B2 (en) 2017-06-16 2023-01-03 Life Technologies Corporation Control nucleic acids, and compositions, kits, and uses thereof
EP3642362A1 (en) 2017-06-20 2020-04-29 Illumina, Inc. Methods and compositions for addressing inefficiencies in amplification reactions
GB201711219D0 (en) 2017-07-12 2017-08-23 Illumina Cambridge Ltd Short pendant arm linkers for nucleotides in sequencing applications
EP3655153B1 (en) 2017-07-18 2020-09-16 Omniome, Inc. Method of chemically modifying plastic surfaces
TWI829642B (zh) 2017-07-24 2024-01-21 美商寬騰矽公司 高強度經標記之反應物組合物及用於定序之方法
US20200202977A1 (en) 2017-07-31 2020-06-25 Illumina, Inc. Sequencing system with multiplexed biological sample aggregation
DK3663407T3 (da) 2017-08-01 2023-04-03 Mgi Tech Co Ltd Fremgangsmåde til nukleinsyresekvensering
EP3545106B1 (en) 2017-08-01 2022-01-19 Helitec Limited Methods of enriching and determining target nucleotide sequences
US11352668B2 (en) 2017-08-01 2022-06-07 Illumina, Inc. Spatial indexing of genetic material and library preparation using hydrogel beads and flow cells
SG11201911871TA (en) 2017-08-01 2020-01-30 Illumina Inc Hydrogel beads for nucleotide sequencing
EP3665274A1 (en) 2017-08-07 2020-06-17 DNA Script Variants of family a dna polymerase and uses thereof
CA3072136A1 (en) 2017-08-15 2019-02-21 Omniome, Inc. Scanning apparatus and methods useful for detection of chemical and biological analytes
US11447818B2 (en) 2017-09-15 2022-09-20 Illumina, Inc. Universal short adapters with variable length non-random unique molecular identifiers
EP3684955A1 (en) 2017-09-20 2020-07-29 Regeneron Pharmaceuticals, Inc. Immunotherapy methods for patients whose tumors carry a high passenger gene mutation burden
NZ759804A (en) 2017-10-16 2022-04-29 Illumina Inc Deep learning-based techniques for training deep convolutional neural networks
AU2018350905B2 (en) 2017-10-16 2021-12-16 Illumina, Inc. Deep learning-based splice site classification
CA3079411C (en) 2017-10-19 2023-12-05 Omniome, Inc. Simultaneous background reduction and complex stabilization in binding assay workflows
EP4289996A3 (en) 2017-11-06 2024-01-17 Illumina Inc. Nucleic acid indexing techniques
NZ759171A (en) 2017-11-16 2022-05-27 Illumina Inc Systems and methods for determining microsatellite instability
WO2019108972A1 (en) 2017-11-30 2019-06-06 Illumina, Inc. Validation methods and systems for sequence variant calls
CN111741967A (zh) 2017-11-30 2020-10-02 深圳市真迈生物科技有限公司 核苷类似物、制备方法及应用
JP6868541B2 (ja) * 2017-12-05 2021-05-12 イルミナ ケンブリッジ リミテッド 修飾ヌクレオシドまたは修飾ヌクレオチド
WO2019136376A1 (en) 2018-01-08 2019-07-11 Illumina, Inc. High-throughput sequencing with semiconductor-based detection
US10752887B2 (en) 2018-01-08 2020-08-25 Dna Script Variants of terminal deoxynucleotidyl transferase and uses thereof
JP7104072B2 (ja) 2018-01-08 2022-07-20 イルミナ インコーポレイテッド 半導体ベースの検出を用いた高スループットシーケンシングのためのシステムおよびデバイス
US10435676B2 (en) 2018-01-08 2019-10-08 Dna Script Variants of terminal deoxynucleotidyl transferase and uses thereof
JP6862581B2 (ja) 2018-01-15 2021-04-21 イルミナ インコーポレイテッド 深層学習ベースのバリアント分類器
US10844430B2 (en) 2018-01-24 2020-11-24 Qiagen Sciences, Llc DNA sequencing reaction additive
RU2020128413A (ru) 2018-01-29 2022-04-01 Зе Броад Институт, Инк. Диагностика на основе эффекторной системы crispr
JP2021512648A (ja) 2018-02-06 2021-05-20 オムニオム・インコーポレイテッド 核酸プライマー伸長のための組成物および技術
CN111094589A (zh) 2018-02-13 2020-05-01 伊鲁米纳公司 使用水凝胶珠的dna测序
CN111164222A (zh) 2018-02-16 2020-05-15 伊鲁米纳公司 带有电荷标签的核苷酸及其使用方法
AU2019248635B2 (en) 2018-04-02 2022-01-27 Illumina, Inc. Compositions and methods for making controls for sequence-based genetic testing
WO2019200338A1 (en) 2018-04-12 2019-10-17 Illumina, Inc. Variant classifier based on deep neural networks
EP3782158A1 (en) 2018-04-19 2021-02-24 Omniome, Inc. Improving accuracy of base calls in nucleic acid sequencing methods
RU2750567C2 (ru) 2018-04-20 2021-06-29 Иллумина, Инк. Способы инкапсулирования одиночных клеток, инкапсулированные клетки и способы их применения
US20210189460A1 (en) 2018-04-25 2021-06-24 Qiagen Sciences Llc Sequential paired-end sequencing
CN112567047A (zh) 2018-04-26 2021-03-26 欧姆尼欧美公司 用于稳定核酸-核苷酸-聚合酶复合物的方法和组合物
AU2019271028A1 (en) 2018-05-14 2020-12-03 Nanostring Technologies, Inc. Chemical compositions and methods of using same
FI3794012T3 (fi) 2018-05-15 2024-01-08 Illumina Inc Koostumuksia ja menetelmiä pintaan sitoutuneiden oligonukleotidien kemiallista pilkkomista ja suojauksen poistoa varten
CN112534063A (zh) 2018-05-22 2021-03-19 安序源有限公司 用于核酸测序的方法、系统和组合物
EP3802880A1 (en) 2018-05-25 2021-04-14 Illumina, Inc. Circulating rna signatures specific to preeclampsia
US11821023B2 (en) 2018-05-30 2023-11-21 Generation Biotech, Llc Method for capturing long polynucleotides for sequencing specific gnomic regions
US11339428B2 (en) 2018-05-31 2022-05-24 Pacific Biosciences Of California, Inc. Increased signal to noise in nucleic acid sequencing
US11180794B2 (en) 2018-05-31 2021-11-23 Omniome, Inc. Methods and compositions for capping nucleic acids
EP4269618A3 (en) 2018-06-04 2024-01-10 Illumina, Inc. Methods of making high-throughput single-cell transcriptome libraries
MX2020013788A (es) 2018-06-22 2021-03-02 Quantum Si Inc Fotodetector integrado con deposito de almacenamiento de carga con tiempo de deteccion variado.
US20200251183A1 (en) 2018-07-11 2020-08-06 Illumina, Inc. Deep Learning-Based Framework for Identifying Sequence Patterns that Cause Sequence-Specific Errors (SSEs)
JP7430701B2 (ja) 2018-07-23 2024-02-13 ディーエヌエー スクリプト 核酸鎖の大規模並列酵素合成
AU2019312152A1 (en) 2018-07-24 2021-02-18 Pacific Biosciences Of California, Inc. Serial formation of ternary complex species
WO2020022891A2 (en) 2018-07-26 2020-01-30 Stichting Vumc Biomarkers for atrial fibrillation
CN112770776A (zh) 2018-07-30 2021-05-07 瑞德库尔有限责任公司 用于样品处理或分析的方法和系统
CN113286884A (zh) 2018-08-07 2021-08-20 博德研究所 新型cas12b酶和系统
KR20210043634A (ko) 2018-08-15 2021-04-21 일루미나, 인코포레이티드 라이브러리 인리치먼트를 개선하기 위한 조성물 및 방법
US20210317524A1 (en) 2018-08-28 2021-10-14 10X Genomics, Inc. Resolving spatial arrays
US11519033B2 (en) 2018-08-28 2022-12-06 10X Genomics, Inc. Method for transposase-mediated spatial tagging and analyzing genomic DNA in a biological sample
CN113366117A (zh) 2018-08-28 2021-09-07 10X基因组学股份有限公司 用于生物样品中转座酶介导的空间标记和分析基因组dna的方法
WO2020056044A1 (en) 2018-09-11 2020-03-19 Singular Genomics Systems, Inc. Modified archaeal family b polymerases
US10731141B2 (en) 2018-09-17 2020-08-04 Omniome, Inc. Engineered polymerases for improved sequencing
EP3856753A4 (en) * 2018-09-28 2022-11-16 Centrillion Technology Holdings Corporation DISULFIDE CROSSLINKED REVERSIBLE TERMINATORS
US20210396756A1 (en) 2018-10-03 2021-12-23 The Broad Institute, Inc. Crispr effector system based diagnostics for hemorrhagic fever detection
NZ759665A (en) 2018-10-15 2022-07-01 Illumina Inc Deep learning-based techniques for pre-training deep convolutional neural networks
US10738072B1 (en) 2018-10-25 2020-08-11 Singular Genomics Systems, Inc. Nucleotide analogues
US11085036B2 (en) 2018-10-26 2021-08-10 Illumina, Inc. Modulating polymer beads for DNA processing
SG11202012493WA (en) 2018-10-31 2021-01-28 Illumina Inc Polymerases, compositions, and methods of use
JP2022513031A (ja) 2018-11-14 2022-02-07 ディーエヌエー スクリプト 末端デオキシヌクレオチジルトランスフェラーゼバリアントおよびその使用
US20240011086A1 (en) 2018-11-15 2024-01-11 Omniome, Inc. Electronic detection of nucleic acid structure
NL2022043B1 (en) 2018-11-21 2020-06-03 Akershus Univ Hf Tagmentation-Associated Multiplex PCR Enrichment Sequencing
SG11202102700TA (en) 2018-11-30 2021-04-29 Illumina Inc Analysis of multiple analytes using a single assay
GB2578528B (en) 2018-12-04 2021-02-24 Omniome Inc Mixed-phase fluids for nucleic acid sequencing and other analytical assays
CN112639090A (zh) 2018-12-05 2021-04-09 亿明达股份有限公司 聚合酶、组合物及使用方法
CA3103736A1 (en) 2018-12-05 2020-06-11 Illumina Cambridge Limited Methods and compositions for cluster generation by bridge amplification
WO2020123309A1 (en) 2018-12-10 2020-06-18 10X Genomics, Inc. Resolving spatial arrays by proximity-based deconvolution
SG11202105441WA (en) 2018-12-13 2021-06-29 Dna Script Direct oligonucleotide synthesis on cells and biomolecules
GB201820300D0 (en) 2018-12-13 2019-01-30 10X Genomics Inc Method for spatial tagging and analysing genomic DNA in a biological specimen
GB201820341D0 (en) 2018-12-13 2019-01-30 10X Genomics Inc Method for transposase-mediated spatial tagging and analysing genomic DNA in a biological specimen
CA3103520A1 (en) 2018-12-14 2020-06-18 Illumina Cambridge Limited Decreasing phasing with unlabeled nucleotides during sequencing
JP2022512265A (ja) 2018-12-17 2022-02-03 イルミナ ケンブリッジ リミテッド シーケンシング用プライマーオリゴヌクレオチド
KR102594019B1 (ko) 2018-12-17 2023-10-26 일루미나 케임브리지 리미티드 폴리뉴클레오타이드 서열분석에 사용하기 위한 조성물
CA3103633A1 (en) 2018-12-18 2020-06-25 Illumina Cambridge Limited Methods and compositions for paired end sequencing using a single surface primer
PL3899037T3 (pl) 2018-12-19 2024-04-08 Illumina, Inc. Sposoby poprawy priorytetu klonalności klastrów polinukleotydowych
EP3899032A2 (en) 2018-12-20 2021-10-27 Omniome, Inc. Temperature control for analysis of nucleic acids and other analytes
US11293061B2 (en) 2018-12-26 2022-04-05 Illumina Cambridge Limited Sequencing methods using nucleotides with 3′ AOM blocking group
US11473136B2 (en) 2019-01-03 2022-10-18 Ncan Genomics, Inc. Linked target capture
EP3906317A1 (en) 2019-01-03 2021-11-10 DNA Script One pot synthesis of sets of oligonucleotides
US11649485B2 (en) 2019-01-06 2023-05-16 10X Genomics, Inc. Generating capture probes for spatial analysis
US11926867B2 (en) 2019-01-06 2024-03-12 10X Genomics, Inc. Generating capture probes for spatial analysis
JP2022516684A (ja) * 2019-01-08 2022-03-01 シンギュラー・ゲノミクス・システムズ・インコーポレイテッド ヌクレオチドの開裂可能なリンカーおよびその使用
CN112930405A (zh) 2019-01-11 2021-06-08 Illumina剑桥有限公司 复合表面结合的转座体复合物
JP2022518056A (ja) 2019-01-23 2022-03-11 クアンタム－エスアイ インコーポレイテッド 高強度標識されたシーケンシング用の反応組成物および方法
EP3921418A4 (en) 2019-02-06 2023-02-08 Singular Genomics Systems, Inc. COMPOSITIONS AND METHODS FOR NUCLEIC ACID SEQUENCE
EP3924500A1 (en) 2019-02-12 2021-12-22 DNA Script Efficient product cleavage in template-free enzymatic synthesis of polynucleotides
EP3924513B1 (en) 2019-02-14 2023-04-12 Pacific Biosciences of California, Inc. Mitigating adverse impacts of detection systems on nucleic acids and other biological analytes
CA3130693A1 (en) 2019-02-19 2020-08-27 Ultima Genomics, Inc. Linkers and methods for optical detection and sequencing
WO2020172444A1 (en) 2019-02-20 2020-08-27 Omniome, Inc. Scanning apparatus and methods for detecting chemical and biological analytes
EP3931354A1 (en) 2019-02-28 2022-01-05 10X Genomics, Inc. Profiling of biological analytes with spatially barcoded oligonucleotide arrays
AU2020232618A1 (en) 2019-03-01 2021-04-08 Illumina, Inc. High-throughput single-nuclei and single-cell libraries and methods of making and of using
CN114127309A (zh) 2019-03-15 2022-03-01 10X基因组学有限公司 使用空间阵列进行单细胞测序的方法
NL2023314B1 (en) 2019-03-21 2020-09-28 Illumina Inc Artificial intelligence-based quality scoring
NL2023316B1 (en) 2019-03-21 2020-09-28 Illumina Inc Artificial intelligence-based sequencing
NL2023310B1 (en) 2019-03-21 2020-09-28 Illumina Inc Training data generation for artificial intelligence-based sequencing
US11676685B2 (en) 2019-03-21 2023-06-13 Illumina, Inc. Artificial intelligence-based quality scoring
US11210554B2 (en) 2019-03-21 2021-12-28 Illumina, Inc. Artificial intelligence-based generation of sequencing metadata
NL2023311B9 (en) 2019-03-21 2021-03-12 Illumina Inc Artificial intelligence-based generation of sequencing metadata
WO2020191387A1 (en) 2019-03-21 2020-09-24 Illumina, Inc. Artificial intelligence-based base calling
NL2023312B1 (en) 2019-03-21 2020-09-28 Illumina Inc Artificial intelligence-based base calling
WO2020198071A1 (en) 2019-03-22 2020-10-01 10X Genomics, Inc. Three-dimensional spatial analysis
US11609208B2 (en) 2019-04-12 2023-03-21 Western Digital Technologies, Inc. Devices and methods for molecule detection based on thermal stabilities of magnetic nanoparticles
US11738336B2 (en) 2019-04-12 2023-08-29 Western Digital Technologies, Inc. Spin torque oscillator (STO) sensors used in nucleic acid sequencing arrays and detection schemes for nucleic acid sequencing
US11112468B2 (en) 2019-04-12 2021-09-07 Western Digital Technologies, Inc. Magnetoresistive sensor array for molecule detection and related detection schemes
US11579217B2 (en) 2019-04-12 2023-02-14 Western Digital Technologies, Inc. Devices and methods for frequency- and phase-based detection of magnetically-labeled molecules using spin torque oscillator (STO) sensors
US11327073B2 (en) 2019-04-12 2022-05-10 Western Digital Technologies, Inc. Thermal sensor array for molecule detection and related detection schemes
CN113905822B (zh) 2019-04-12 2023-09-22 西部数据技术公司 使用磁传感器阵列边合成边核酸测序
US11593649B2 (en) 2019-05-16 2023-02-28 Illumina, Inc. Base calling using convolutions
WO2020243579A1 (en) 2019-05-30 2020-12-03 10X Genomics, Inc. Methods of detecting spatial heterogeneity of a biological sample
US11644406B2 (en) 2019-06-11 2023-05-09 Pacific Biosciences Of California, Inc. Calibrated focus sensing
AU2020312787A1 (en) 2019-07-12 2021-06-17 Illumina Cambridge Limited Nucleic acid library preparation using electrophoresis
BR112021012755A2 (pt) 2019-07-12 2022-04-26 Illumina Cambridge Ltd Composições e métodos para preparar bibliotecas de sequenciamento de ácidos nucleicos usando crispr/cas9 imobilizadas em um suporte sólido
US11377655B2 (en) 2019-07-16 2022-07-05 Pacific Biosciences Of California, Inc. Synthetic nucleic acids having non-natural structures
US10656368B1 (en) 2019-07-24 2020-05-19 Omniome, Inc. Method and system for biological imaging using a wide field objective lens
WO2021018919A1 (en) 2019-07-30 2021-02-04 Dna Script Template-free enzymatic synthesis of polynucleotides using poly(a) and poly(u) polymerases
CN114430778A (zh) 2019-08-01 2022-05-03 Dna斯克瑞普特公司 增加多核苷酸的无模板酶促合成中长序列的产量
US20220315970A1 (en) 2019-09-09 2022-10-06 Dna Script Template-Free Enzymatic Polynucleotide Synthesis Using Photocleavable Linkages
US11180520B2 (en) 2019-09-10 2021-11-23 Omniome, Inc. Reversible modifications of nucleotides
US20220290245A1 (en) 2019-09-11 2022-09-15 The United States Of America, As Represented By The Secretary, Department Of Health And Human Servic Cancer detection and classification
US11512295B2 (en) 2019-09-12 2022-11-29 Singular Genomics Systems, Inc. Modified thermoccocus polymerases
US11208682B2 (en) 2019-09-13 2021-12-28 Western Digital Technologies, Inc. Enhanced optical detection for nucleic acid sequencing using thermally-dependent fluorophore tags
US11287422B2 (en) 2019-09-23 2022-03-29 Element Biosciences, Inc. Multivalent binding composition for nucleic acid analysis
EP4034669A1 (en) 2019-09-23 2022-08-03 DNA Script Increasing long-sequence yields in template-free enzymatic synthesis of polynucleotides
CN117036248A (zh) 2019-10-01 2023-11-10 10X基因组学有限公司 用于识别组织样品中的形态学模式的系统和方法
WO2021076152A1 (en) 2019-10-18 2021-04-22 Omniome, Inc. Methods and compositions for capping nucleic acids
WO2021092433A2 (en) 2019-11-08 2021-05-14 10X Genomics, Inc. Enhancing specificity of analyte binding
US20210139867A1 (en) 2019-11-08 2021-05-13 Omniome, Inc. Engineered polymerases for improved sequencing by binding
EP4055185A1 (en) 2019-11-08 2022-09-14 10X Genomics, Inc. Spatially-tagged analyte capture agents for analyte multiplexing
US20220411840A1 (en) 2019-11-13 2022-12-29 Dna Script High Efficiency Template-Free Enzymatic Synthesis of Polynucleotides
AU2020388573A1 (en) 2019-11-21 2022-06-23 10X Genomics, Inc, Spatial analysis of analytes
US11747329B2 (en) 2019-11-22 2023-09-05 Western Digital Technologies, Inc. Magnetic gradient concentrator/reluctance detector for molecule detection
CN113677807A (zh) 2019-11-22 2021-11-19 因美纳有限公司 先兆子痫特异性的循环rna标记
DE202019106694U1 (de) 2019-12-02 2020-03-19 Omniome, Inc. System zur Sequenzierung von Nukleinsäuren in Fluidschaum
DE202019106695U1 (de) 2019-12-02 2020-03-19 Omniome, Inc. System zur Sequenzierung von Nukleinsäuren in Fluidschaum
US20210172012A1 (en) 2019-12-04 2021-06-10 Illumina, Inc. Preparation of dna sequencing libraries for detection of dna pathogens in plasma
WO2021118349A1 (en) 2019-12-10 2021-06-17 Prinses Máxima Centrum Voor Kinderoncologie B.V. Methods of typing germ cell tumors
CN114787346A (zh) 2019-12-12 2022-07-22 Dna斯克瑞普特公司 用于多核苷酸无模板酶学合成的嵌合末端脱氧核苷酸转移酶
US20230203553A1 (en) 2019-12-16 2023-06-29 Dna Script Template-Free Enzymatic Polynucleotide Synthesis Using Dismutationless Terminal Deoxynucleotidyl Transferase Variants
JP2023506891A (ja) 2019-12-18 2023-02-20 エフ． ホフマン－ラ ロシュ エージー． 連続的な標識スキームを使用した合成による配列決定方法
CN114008199A (zh) 2019-12-19 2022-02-01 伊路敏纳公司 高通量单细胞文库及其制备和使用方法
CN114885610A (zh) 2019-12-23 2022-08-09 10X基因组学有限公司 使用rna模板化连接进行空间分析的方法
US11225688B2 (en) 2019-12-23 2022-01-18 Singular Genomics Systems, Inc. Methods for long read sequencing
US11747262B2 (en) 2019-12-23 2023-09-05 Singular Genomics Systems, Inc. Flow cell carrier and methods of use
US11498078B2 (en) 2019-12-23 2022-11-15 Singular Genomics Systems, Inc. Flow cell receiver and methods of use
AU2020416718A1 (en) 2019-12-31 2022-07-07 Singular Genomics Systems, Inc. Polynucleotide barcodes for long read sequencing
US11702693B2 (en) 2020-01-21 2023-07-18 10X Genomics, Inc. Methods for printing cells and generating arrays of barcoded cells
US11732299B2 (en) 2020-01-21 2023-08-22 10X Genomics, Inc. Spatial assays with perturbed cells
CN115362264A (zh) 2020-01-29 2022-11-18 10X基因组学有限公司 用于分析物检测的组合物和方法
US11821035B1 (en) 2020-01-29 2023-11-21 10X Genomics, Inc. Compositions and methods of making gene expression libraries
US11898205B2 (en) 2020-02-03 2024-02-13 10X Genomics, Inc. Increasing capture efficiency of spatial assays
US20230054204A1 (en) 2020-02-04 2023-02-23 Pacific Biosciences Of California, Inc. Flow cells and methods for their manufacture and use
US11732300B2 (en) 2020-02-05 2023-08-22 10X Genomics, Inc. Increasing efficiency of spatial analysis in a biological sample
US11835462B2 (en) 2020-02-11 2023-12-05 10X Genomics, Inc. Methods and compositions for partitioning a biological sample
EP4107284A1 (en) 2020-02-17 2022-12-28 10X Genomics, Inc. In situ analysis of chromatin interaction
US11807851B1 (en) 2020-02-18 2023-11-07 Ultima Genomics, Inc. Modified polynucleotides and uses thereof
US11749380B2 (en) 2020-02-20 2023-09-05 Illumina, Inc. Artificial intelligence-based many-to-many base calling
US20210265016A1 (en) 2020-02-20 2021-08-26 Illumina, Inc. Data Compression for Artificial Intelligence-Based Base Calling
US20210265015A1 (en) 2020-02-20 2021-08-26 Illumina, Inc. Hardware Execution and Acceleration of Artificial Intelligence-Based Base Caller
US20210265018A1 (en) 2020-02-20 2021-08-26 Illumina, Inc. Knowledge Distillation and Gradient Pruning-Based Compression of Artificial Intelligence-Based Base Caller
US20210262018A1 (en) 2020-02-21 2021-08-26 10X Genomics, Inc. Methods and compositions for integrated in situ spatial assay
CN115461143A (zh) 2020-02-21 2022-12-09 10X基因组学有限公司 用于样品处理的组合物、方法和系统
US11891654B2 (en) 2020-02-24 2024-02-06 10X Genomics, Inc. Methods of making gene expression libraries
EP4110940B1 (en) 2020-02-25 2024-03-27 DNA Script Method and apparatus for enzymatic synthesis of polynucleotides
US11034942B1 (en) 2020-02-27 2021-06-15 Singular Genomics Systems, Inc. Modified pyrococcus polymerases and uses thereof
US11926863B1 (en) 2020-02-27 2024-03-12 10X Genomics, Inc. Solid state single cell method for analyzing fixed biological cells
CN115516104A (zh) 2020-03-03 2022-12-23 加利福尼亚太平洋生物科学股份有限公司 用于对双链核酸进行测序的方法和组合物
US11768175B1 (en) 2020-03-04 2023-09-26 10X Genomics, Inc. Electrophoretic methods for spatial analysis
WO2021178893A2 (en) 2020-03-06 2021-09-10 Singular Genomics Systems, Inc. Linked paired strand sequencing
US20230340589A1 (en) 2020-03-10 2023-10-26 Western Digital Technologies, Inc. Magnetic sensor arrays for nucleic acid sequencing and methods of making and using them
BR112022019541A2 (pt) 2020-03-30 2022-11-16 Illumina Inc Métodos e composições para preparação de bibliotecas de ácido nucleico
AU2021260014A1 (en) 2020-04-20 2022-11-17 Dna Script Terminal deoxynucleotidyl transferase variants and uses thereof
WO2021216708A1 (en) 2020-04-22 2021-10-28 10X Genomics, Inc. Methods for spatial analysis using targeted rna depletion
WO2021216998A1 (en) 2020-04-24 2021-10-28 Singular Genomics Systems, Inc. Modified nucleotides and uses thereof
US20230330223A1 (en) 2020-04-27 2023-10-19 Agendia N.V. Treatment of her2 negative, mammaprint high risk 2 breast cancer
EP4146822A1 (en) 2020-05-05 2023-03-15 Pacific Biosciences of California, Inc. Compositions and methods for modifying polymerase-nucleic acid complexes
US11188778B1 (en) 2020-05-05 2021-11-30 Illumina, Inc. Equalization-based image processing and spatial crosstalk attenuator
WO2021224677A1 (en) 2020-05-05 2021-11-11 Akershus Universitetssykehus Hf Compositions and methods for characterizing bowel cancer
JP2023524531A (ja) 2020-05-08 2023-06-12 イルミナ インコーポレイテッド ゲノムシーケンシング及び検出手法
CN115916967A (zh) 2020-05-12 2023-04-04 伊鲁米纳公司 使用重组末端脱氧核苷酸转移酶生成具有经修饰的碱基的核酸
WO2021237087A1 (en) 2020-05-22 2021-11-25 10X Genomics, Inc. Spatial analysis to detect sequence variants
EP4153775A1 (en) 2020-05-22 2023-03-29 10X Genomics, Inc. Simultaneous spatio-temporal measurement of gene expression and cellular activity
WO2021242834A1 (en) 2020-05-26 2021-12-02 10X Genomics, Inc. Method for resetting an array
EP4025692A2 (en) 2020-06-02 2022-07-13 10X Genomics, Inc. Nucleic acid library methods
CN116249785A (zh) 2020-06-02 2023-06-09 10X基因组学有限公司 用于抗原-受体的空间转录组学
EP4162074B1 (en) 2020-06-08 2024-04-24 10X Genomics, Inc. Methods of determining a surgical margin and methods of use thereof
WO2021252617A1 (en) 2020-06-09 2021-12-16 Illumina, Inc. Methods for increasing yield of sequencing libraries
EP4165207A1 (en) 2020-06-10 2023-04-19 10X Genomics, Inc. Methods for determining a location of an analyte in a biological sample
EP4165549A1 (en) 2020-06-11 2023-04-19 Nautilus Biotechnology, Inc. Methods and systems for computational decoding of biological, chemical, and physical entities
IL299164A (en) 2020-06-16 2023-02-01 Dna Script Systems, devices and kits for the enzymatic synthesis of polynucleotides
MX2022016492A (es) 2020-06-22 2023-03-06 Illumina Cambridge Ltd Nucleosidos y nucleotidos con grupo de bloqueo de acetal 3'.
WO2021263111A1 (en) 2020-06-25 2021-12-30 10X Genomics, Inc. Spatial analysis of dna methylation
CN115997033A (zh) 2020-06-30 2023-04-21 伊鲁米纳公司 通过合成产生无痕dna的催化控制测序
AU2021300252A1 (en) 2020-07-02 2023-01-05 Illumina, Inc. A method to calibrate nucleic acid library seeding efficiency in flowcells
US11761038B1 (en) 2020-07-06 2023-09-19 10X Genomics, Inc. Methods for identifying a location of an RNA in a biological sample
CN116018412A (zh) 2020-07-08 2023-04-25 Illumina公司 作为转座体载体的小珠
EP4153606A2 (en) 2020-07-13 2023-03-29 Singular Genomics Systems, Inc. Methods of sequencing complementary polynucleotides
WO2022013094A1 (en) 2020-07-15 2022-01-20 Dna Script Massively parallel enzymatic synthesis of polynucleotides
CA3190588A1 (en) 2020-08-06 2022-02-10 Illumina, Inc. Preparation of rna and dna sequencing libraries using bead-linked transposomes
AU2021329302A1 (en) 2020-08-18 2023-02-16 Illumina Cambridge Limited Sequence-specific targeted transposition and selection and sorting of nucleic acids
US20220067489A1 (en) 2020-08-28 2022-03-03 Illumina, Inc. Detecting and Filtering Clusters Based on Artificial Intelligence-Predicted Base Calls
US11200446B1 (en) 2020-08-31 2021-12-14 Element Biosciences, Inc. Single-pass primary analysis
WO2022053610A1 (en) 2020-09-11 2022-03-17 Illumina Cambridge Limited Methods of enriching a target sequence from a sequencing library using hairpin adaptors
WO2022056385A1 (en) 2020-09-14 2022-03-17 Singular Genomics Systems, Inc. Methods and systems for multidimensional imaging
WO2022063835A1 (en) 2020-09-22 2022-03-31 Dna Script Stabilized n-terminally truncated terminal deoxynucleotidyl transferase variants and uses thereof
US11926822B1 (en) 2020-09-23 2024-03-12 10X Genomics, Inc. Three-dimensional spatial analysis
CN116438319A (zh) 2020-10-21 2023-07-14 Illumina公司 包含多个插入物的测序模板以及用于提高测序通量的组合物和方法
WO2022090323A1 (en) 2020-10-29 2022-05-05 Dna Script Enzymatic synthesis of polynucleotide probes
US20220145345A1 (en) 2020-11-11 2022-05-12 Microsoft Technology Licensing, Llc Spatial control of polynucleotide synthesis by strand capping
US11827935B1 (en) 2020-11-19 2023-11-28 10X Genomics, Inc. Methods for spatial analysis using rolling circle amplification and detection probes
US20240044906A1 (en) * 2020-12-01 2024-02-08 Kyushu University, National University Corporation Biomolecule structure detection probe, biomolecule structure detection kit, and method for detecting biomolecule structure
JP2023552015A (ja) 2020-12-02 2023-12-14 イルミナ ソフトウェア， インコーポレイテッド 遺伝子変異を検出するためのシステム及び方法
EP4012046A1 (en) 2020-12-11 2022-06-15 10X Genomics, Inc. Methods and compositions for multimodal in situ analysis
JP2024501232A (ja) 2020-12-21 2024-01-11 シンギュラー・ゲノミクス・システムズ・インコーポレイテッド 多色撮像のためのシステム及び方法
WO2022140028A1 (en) 2020-12-21 2022-06-30 10X Genomics, Inc. Methods, compositions, and systems for capturing probes and/or barcodes
WO2022155331A1 (en) 2021-01-13 2022-07-21 Pacific Biosciences Of California, Inc. Surface structuring with colloidal assembly
US20220228200A1 (en) 2021-01-19 2022-07-21 10X Genomics, Inc. Methods and compositions for internally controlled in situ assays
EP4284945A1 (en) 2021-01-26 2023-12-06 10X Genomics, Inc. Nucleic acid analog probes for in situ analysis
CA3206511A1 (en) 2021-01-29 2022-08-04 Yir-Shyuan WU Methods, compositions and kits to improve seeding efficiency of flow cells with polynucleotides
CA3208854A1 (en) 2021-02-04 2022-08-11 Illumina, Inc. Long indexed-linked read generation on transposome bound beads
WO2022170212A1 (en) 2021-02-08 2022-08-11 Singular Genomics Systems, Inc. Methods and compositions for sequencing complementary polynucleotides
WO2022174054A1 (en) 2021-02-13 2022-08-18 The General Hospital Corporation Methods and compositions for in situ macromolecule detection and uses thereof
US20220282319A1 (en) 2021-03-03 2022-09-08 10X Genomics, Inc. Analyte detection in situ using nucleic acid origami
EP4263868A1 (en) 2021-03-12 2023-10-25 Singular Genomics Systems, Inc. Nanoarrays and methods of use thereof
US11884977B2 (en) 2021-03-12 2024-01-30 Singular Genomics Systems, Inc. Nanoarrays and methods of use thereof
WO2022197752A1 (en) 2021-03-16 2022-09-22 Illumina, Inc. Tile location and/or cycle based weight set selection for base calling
WO2022198068A1 (en) 2021-03-18 2022-09-22 10X Genomics, Inc. Multiplex capture of gene and protein expression from a biological sample
CA3210451A1 (en) 2021-03-22 2022-09-29 Illumina Cambridge Limited Methods for improving nucleic acid cluster clonality
WO2022212280A1 (en) 2021-03-29 2022-10-06 Illumina, Inc. Compositions and methods for assessing dna damage in a library and normalizing amplicon size bias
CA3214278A1 (en) 2021-03-29 2022-10-06 Illumina, Inc Improved methods of library preparation
CA3214584A1 (en) 2021-03-30 2022-10-06 Illumina, Inc. Improved methods of isothermal complementary dna and library preparation
IL307159A (en) 2021-03-31 2023-11-01 Illumina Cambridge Ltd Nucleic acid library sequencing techniques with adapter dimer detection
AU2022248999A1 (en) 2021-03-31 2023-02-02 Illumina, Inc. Artificial intelligence-based base caller with contextual awareness
BR112023019945A2 (pt) 2021-03-31 2023-11-14 Illumina Cambridge Ltd Métodos para a preparação de bibliotecas de sequenciamento por tagmentação direcional com o uso de tecnologia baseada em transposon com identificadores moleculares exclusivos para correção de erros
WO2022207934A1 (en) 2021-04-02 2022-10-06 Dna Script Methods and kits for enzymatic synthesis of g4-prone polynucleotides
WO2022213027A1 (en) 2021-04-02 2022-10-06 Illumina, Inc. Machine-learning model for detecting a bubble within a nucleotide-sample slide for sequencing
US20220336054A1 (en) 2021-04-15 2022-10-20 Illumina, Inc. Deep Convolutional Neural Networks to Predict Variant Pathogenicity using Three-Dimensional (3D) Protein Structures
FR3121936A1 (fr) 2021-04-20 2022-10-21 Naos Institute Of Life Science Methode de criblage d’ingredients et de compositions topiques ecobiologiques, et son utilisation pour realiser un soin cosmetique de restauration du microbiote cutane
WO2022232308A1 (en) 2021-04-27 2022-11-03 Singular Genomics Systems, Inc. High density sequencing and multiplexed priming
EP4330430A2 (en) 2021-04-29 2024-03-06 Illumina, Inc. Amplification techniques for nucleic acid characterization
WO2022235163A1 (en) 2021-05-07 2022-11-10 Agendia N.V. Endocrine treatment of hormone receptor positive breast cancer typed as having a low risk of recurrence
WO2022240764A1 (en) 2021-05-10 2022-11-17 Pacific Biosciences Of California, Inc. Single-molecule seeding and amplification on a surface
CN117858959A (zh) 2021-05-10 2024-04-09 加利福尼亚太平洋生物科学股份有限公司 滚环扩增期间的dna扩增缓冲液补充
WO2022243480A1 (en) 2021-05-20 2022-11-24 Illumina, Inc. Compositions and methods for sequencing by synthesis
WO2022251510A2 (en) 2021-05-28 2022-12-01 Illumina, Inc. Oligo-modified nucleotide analogues for nucleic acid preparation
US20220380838A1 (en) 2021-06-01 2022-12-01 10X Genomics, Inc. Methods and compositions for analyte detection and probe resolution
US20230084407A1 (en) 2021-06-02 2023-03-16 10X Genomics, Inc. Sample analysis using asymmetric circularizable probes
US20220403450A1 (en) 2021-06-03 2022-12-22 Illumina Software, Inc. Systems and methods for sequencing nucleotides using two optical channels
WO2022265994A1 (en) 2021-06-15 2022-12-22 Illumina, Inc. Hydrogel-free surface functionalization for sequencing
US11220707B1 (en) 2021-06-17 2022-01-11 Element Biosciences, Inc. Compositions and methods for pairwise sequencing
US11859241B2 (en) 2021-06-17 2024-01-02 Element Biosciences, Inc. Compositions and methods for pairwise sequencing
CA3223112A1 (en) 2021-06-18 2022-12-22 Mark AMBROSO Engineered polymerases
WO2022272260A1 (en) 2021-06-23 2022-12-29 Illumina, Inc. Compositions, methods, kits, cartridges, and systems for sequencing reagents
CA3224393A1 (en) 2021-06-29 2023-01-05 Mitchell A BEKRITSKY Machine-learning model for generating confidence classifications for genomic coordinates
KR20240022490A (ko) 2021-06-29 2024-02-20 일루미나, 인코포레이티드 뉴클레오티드 염기 호출 및 염기 호출 품질을 결정하기 위한 신호-대-잡음비 메트릭
WO2023278184A1 (en) 2021-06-29 2023-01-05 Illumina, Inc. Methods and systems to correct crosstalk in illumination emitted from reaction sites
CA3224382A1 (en) 2021-06-29 2023-01-05 Amirali Kia Self-learned base caller, trained using oligo sequences
US20230005253A1 (en) 2021-07-01 2023-01-05 Illumina, Inc. Efficient artificial intelligence-based base calling of index sequences
US20230027409A1 (en) 2021-07-13 2023-01-26 Illumina, Inc. Methods and systems for real time extraction of crosstalk in illumination emitted from reaction sites
US20230026886A1 (en) 2021-07-13 2023-01-26 10X Genomics, Inc. Methods for preparing polymerized matrix with controllable thickness
US11455487B1 (en) 2021-10-26 2022-09-27 Illumina Software, Inc. Intensity extraction and crosstalk attenuation using interpolation and adaptation for base calling
WO2023003757A1 (en) 2021-07-19 2023-01-26 Illumina Software, Inc. Intensity extraction with interpolation and adaptation for base calling
US20230021577A1 (en) 2021-07-23 2023-01-26 Illumina Software, Inc. Machine-learning model for recalibrating nucleotide-base calls
WO2023004357A1 (en) 2021-07-23 2023-01-26 Illumina, Inc. Methods for preparing substrate surface for dna sequencing
KR20240037882A (ko) 2021-07-28 2024-03-22 일루미나, 인코포레이티드 염기 호출 시스템의 품질 점수 보정
WO2023014741A1 (en) 2021-08-03 2023-02-09 Illumina Software, Inc. Base calling using multiple base caller models
US20230057571A1 (en) 2021-08-03 2023-02-23 10X Genomics, Inc. Nucleic acid concatemers and methods for stabilizing and/or compacting the same
US20230047225A1 (en) 2021-08-14 2023-02-16 Illumina, Inc. Polymerases, compositions, and methods of use
EP4326898A1 (en) 2021-08-16 2024-02-28 10X Genomics, Inc. Probes comprising a split barcode region and methods of use
AU2022331421A1 (en) 2021-08-17 2024-01-04 Illumina, Inc. Methods and compositions for identifying methylated cytosines
WO2023034489A1 (en) 2021-09-01 2023-03-09 10X Genomics, Inc. Methods, compositions, and kits for blocking a capture probe on a spatial array
WO2023034079A1 (en) 2021-09-01 2023-03-09 Illumina Software, Inc. Amplitude modulation for accelerated base calling
WO2023034920A2 (en) 2021-09-03 2023-03-09 Singular Genomics Systems, Inc. Amplification oligonucleotides
WO2023044229A1 (en) 2021-09-17 2023-03-23 Illumina, Inc. Automatically identifying failure sources in nucleotide sequencing from base-call-error patterns
CN117546243A (zh) 2021-09-21 2024-02-09 因美纳有限公司 使用推算的单倍型的图参考基因组和碱基检出方法
WO2023049212A2 (en) 2021-09-22 2023-03-30 Illumina, Inc. State-based base calling
US20230096386A1 (en) 2021-09-30 2023-03-30 Illumina Cambridge Limited Polynucleotide sequencing
WO2023056328A2 (en) 2021-09-30 2023-04-06 Illumina, Inc. Solid supports and methods for depleting and/or enriching library fragments prepared from biosamples
WO2023064181A1 (en) 2021-10-11 2023-04-20 Nautilus Biotechnology, Inc. Highly multiplexable analysis of proteins and proteomes
WO2023069927A1 (en) 2021-10-20 2023-04-27 Illumina, Inc. Methods for capturing library dna for sequencing
WO2023081485A1 (en) 2021-11-08 2023-05-11 Pacific Biosciences Of California, Inc. Stepwise sequencing of a polynucleotide with a homogenous reaction mixture
WO2023083997A2 (en) 2021-11-10 2023-05-19 Dna Script Novel terminal deoxynucleotidyl
WO2023083999A2 (en) 2021-11-10 2023-05-19 Dna Script Novel terminal deoxynucleotidyl transferase (tdt) variants
WO2023085932A1 (en) 2021-11-10 2023-05-19 Omnigen B.V. Prediction of response following folfirinox treatment in cancer patients
EP4305195A2 (en) 2021-12-01 2024-01-17 10X Genomics, Inc. Methods, compositions, and systems for improved in situ detection of analytes and spatial analysis
CN117581303A (zh) 2021-12-02 2024-02-20 因美纳有限公司 产生用于确定核苷酸碱基检出的簇特异性信号校正
US20230183799A1 (en) 2021-12-10 2023-06-15 Illumina, Inc. Parallel sample and index sequencing
WO2023119164A1 (en) 2021-12-21 2023-06-29 Illumina Cambridge Limited Wax-microsphere matrix compositions and methods of making and using the same
US20230215515A1 (en) 2021-12-23 2023-07-06 Illumina Software, Inc. Facilitating secure execution of external workflows for genomic sequencing diagnostics
WO2023122363A1 (en) 2021-12-23 2023-06-29 Illumina Software, Inc. Dynamic graphical status summaries for nucelotide sequencing
WO2023129898A2 (en) 2021-12-27 2023-07-06 10X Genomics, Inc. Methods and compositions for rolling circle amplification
US20230207050A1 (en) 2021-12-28 2023-06-29 Illumina Software, Inc. Machine learning model for recalibrating nucleotide base calls corresponding to target variants
WO2023129764A1 (en) 2021-12-29 2023-07-06 Illumina Software, Inc. Automatically switching variant analysis model versions for genomic analysis applications
US20230212667A1 (en) 2021-12-29 2023-07-06 Illumina Cambridge Limited Methods of nucleic acid sequencing using surface-bound primers
CA3223362A1 (en) 2022-01-20 2023-07-27 Xiaolin Wu Methods of detecting methylcytosine and hydroxymethylcytosine by sequencing
US20230279475A1 (en) 2022-01-21 2023-09-07 10X Genomics, Inc. Multiple readout signals for analyzing a sample
WO2023164570A1 (en) 2022-02-23 2023-08-31 Insitro, Inc. Pooled optical screening and transcriptional measurements of cells comprising barcoded genetic perturbations
WO2023164660A1 (en) 2022-02-25 2023-08-31 Illumina, Inc. Calibration sequences for nucelotide sequencing
WO2023164492A1 (en) 2022-02-25 2023-08-31 Illumina, Inc. Machine-learning models for detecting and adjusting values for nucleotide methylation levels
US11795505B2 (en) 2022-03-10 2023-10-24 Singular Genomics Systems, Inc. Nucleic acid delivery scaffolds
WO2023175026A1 (en) 2022-03-15 2023-09-21 Illumina, Inc. Methods of determining sequence information
WO2023183937A1 (en) 2022-03-25 2023-09-28 Illumina, Inc. Sequence-to-sequence base calling
WO2023192917A1 (en) 2022-03-29 2023-10-05 Nautilus Subsidiary, Inc. Integrated arrays for single-analyte processes
WO2023192616A1 (en) 2022-04-01 2023-10-05 10X Genomics, Inc. Compositions and methods for targeted masking of autofluorescence
US20230323427A1 (en) 2022-04-06 2023-10-12 10X Genomics, Inc. Methods and compositions for multiplex cell analysis
WO2023196572A1 (en) 2022-04-07 2023-10-12 Illumina Singapore Pte. Ltd. Altered cytidine deaminases and methods of use
WO2023196528A1 (en) 2022-04-08 2023-10-12 Illumina, Inc. Aptamer dynamic range compression and detection techniques
WO2023212490A1 (en) 2022-04-25 2023-11-02 Nautilus Subsidiary, Inc. Systems and methods for assessing and improving the quality of multiplex molecular assays
US20230340571A1 (en) 2022-04-26 2023-10-26 Illumina, Inc. Machine-learning models for selecting oligonucleotide probes for array technologies
WO2023209606A1 (en) 2022-04-29 2023-11-02 Illumina Cambridge Limited Methods and systems for encapsulating lyophilised microspheres
US20240002902A1 (en) 2022-05-06 2024-01-04 10X Genomics, Inc. Analysis of antigen and antigen receptor interactions
WO2023220627A1 (en) 2022-05-10 2023-11-16 Illumina Software, Inc. Adaptive neural network for nucelotide sequencing
WO2023225095A1 (en) 2022-05-18 2023-11-23 Illumina Cambridge Limited Preparation of size-controlled nucleic acid fragments
WO2023224487A1 (en) 2022-05-19 2023-11-23 Agendia N.V. Prediction of response to immune therapy in breast cancer patients
WO2023224488A1 (en) 2022-05-19 2023-11-23 Agendia N.V. Dna repair signature and prediction of response following cancer therapy
US20230392207A1 (en) 2022-06-03 2023-12-07 Illumina, Inc. Circulating rna biomarkers for preeclampsia
WO2023245190A1 (en) 2022-06-17 2023-12-21 10X Genomics, Inc. Catalytic de-crosslinking of samples for in situ analysis
US20230420080A1 (en) 2022-06-24 2023-12-28 Illumina Software, Inc. Split-read alignment by intelligently identifying and scoring candidate split groups
US20230420075A1 (en) 2022-06-27 2023-12-28 Illumina Software, Inc. Accelerators for a genotype imputation model
US20230420082A1 (en) 2022-06-27 2023-12-28 Illumina Software, Inc. Generating and implementing a structural variation graph genome
WO2024006705A1 (en) 2022-06-27 2024-01-04 Illumina Software, Inc. Improved human leukocyte antigen (hla) genotyping
WO2024015962A1 (en) 2022-07-15 2024-01-18 Pacific Biosciences Of California, Inc. Blocked asymmetric hairpin adaptors
WO2024026356A1 (en) 2022-07-26 2024-02-01 Illumina, Inc. Rapid single-cell multiomics processing using an executable file
WO2024036304A1 (en) 2022-08-12 2024-02-15 10X Genomics, Inc. Puma1 polymerases and uses thereof
US20240084373A1 (en) 2022-08-16 2024-03-14 10X Genomics, Inc. Ap50 polymerases and uses thereof
WO2024039516A1 (en) 2022-08-19 2024-02-22 Illumina, Inc. Third dna base pair site-specific dna detection
WO2024059655A1 (en) 2022-09-15 2024-03-21 Nautilus Subsidiary, Inc. Characterizing accessibility of macromolecule structures
WO2024059852A1 (en) 2022-09-16 2024-03-21 Illumina, Inc. Cluster segmentation and conditional base calling
WO2024073516A1 (en) 2022-09-29 2024-04-04 Illumina, Inc. A target-variant-reference panel for imputing target variants
US20240120027A1 (en) 2022-09-30 2024-04-11 Illumina, Inc. Machine-learning model for refining structural variant calls
WO2024073043A1 (en) 2022-09-30 2024-04-04 Illumina, Inc. Methods of using cpg binding proteins in mapping modified cytosine nucleotides
WO2024068971A1 (en) 2022-09-30 2024-04-04 Illumina, Inc. Polymerases, compositions, and methods of use
WO2024069581A1 (en) 2022-09-30 2024-04-04 Illumina Singapore Pte. Ltd. Helicase-cytidine deaminase complexes and methods of use
WO2024073047A1 (en) 2022-09-30 2024-04-04 Illumina, Inc. Cytidine deaminases and methods of use in mapping modified cytosine nucleotides
WO2024077096A1 (en) 2022-10-05 2024-04-11 Illumina, Inc. Integrating variant calls from multiple sequencing pipelines utilizing a machine learning architecture
WO2024077152A1 (en) 2022-10-06 2024-04-11 Illumina, Inc. Probes for depleting abundant small noncoding rna
WO2024077162A2 (en) 2022-10-06 2024-04-11 Illumina, Inc. Probes for improving coronavirus sample surveillance
WO2024077202A2 (en) 2022-10-06 2024-04-11 Illumina, Inc. Probes for improving environmental sample surveillance

Family Cites Families (90)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4711955A (en) * 1981-04-17 1987-12-08 Yale University Modified nucleotides and methods of preparing and using same
US5175269A (en) 1984-01-30 1992-12-29 Enzo Diagnostics, Inc. Compound and detectable molecules having an oligo- or polynucleotide with modifiable reactive group
US5118605A (en) * 1984-10-16 1992-06-02 Chiron Corporation Polynucleotide determination with selectable cleavage sites
US4824775A (en) * 1985-01-03 1989-04-25 Molecular Diagnostics, Inc. Cells labeled with multiple Fluorophores bound to a nucleic acid carrier
US4772691A (en) 1985-06-05 1988-09-20 The Medical College Of Wisconsin, Inc. Chemically cleavable nucleotides
US4863849A (en) * 1985-07-18 1989-09-05 New York Medical College Automatable process for sequencing nucleotide
US5151507A (en) 1986-07-02 1992-09-29 E. I. Du Pont De Nemours And Company Alkynylamino-nucleotides
SE8801070D0 (sv) 1988-03-23 1988-03-23 Pharmacia Ab Method for immobilizing a dna sequence on a solid support
US5174962A (en) * 1988-06-20 1992-12-29 Genomyx, Inc. Apparatus for determining DNA sequences by mass spectrometry
GB8910880D0 (en) 1989-05-11 1989-06-28 Amersham Int Plc Sequencing method
US5547839A (en) * 1989-06-07 1996-08-20 Affymax Technologies N.V. Sequencing of surface immobilized polymers utilizing microflourescence detection
JPH07111484B2 (ja) * 1989-06-26 1995-11-29 松下電器産業株式会社 プラスチック製光学部品の反射防止膜とその形成方法
US5302509A (en) 1989-08-14 1994-04-12 Beckman Instruments, Inc. Method for sequencing polynucleotides
EP0450060A1 (en) * 1989-10-26 1991-10-09 Sri International Dna sequencing
AU2674092A (en) 1991-09-09 1993-04-05 Baylor College Of Medicine Method and device for rapid dna or rna sequencing determination by a base addition sequencing scheme
DE4141178A1 (de) 1991-12-13 1993-06-17 Europ Lab Molekularbiolog Verfahren zur sequenzierung von nukleinsaeuren
GB9208733D0 (en) 1992-04-22 1992-06-10 Medical Res Council Dna sequencing method
GB9210176D0 (en) * 1992-05-12 1992-06-24 Cemu Bioteknik Ab Chemical method
US5436143A (en) 1992-12-23 1995-07-25 Hyman; Edward D. Method for enzymatic synthesis of oligonucleotides
US5516664A (en) 1992-12-23 1996-05-14 Hyman; Edward D. Enzymatic synthesis of repeat regions of oligonucleotides
US6074823A (en) * 1993-03-19 2000-06-13 Sequenom, Inc. DNA sequencing by mass spectrometry via exonuclease degradation
FR2703052B1 (fr) 1993-03-26 1995-06-02 Pasteur Institut Nouvelle méthode de séquençage d'acides nucléiques.
GB9315847D0 (en) * 1993-07-30 1993-09-15 Isis Innovation Tag reagent and assay method
US5547859A (en) * 1993-08-02 1996-08-20 Goodman; Myron F. Chain-terminating nucleotides for DNA sequencing methods
US5849542A (en) * 1993-11-17 1998-12-15 Amersham Pharmacia Biotech Uk Limited Primer extension mass spectroscopy nucleic acid sequencing method
US6232465B1 (en) * 1994-09-02 2001-05-15 Andrew C. Hiatt Compositions for enzyme catalyzed template-independent creation of phosphodiester bonds using protected nucleotides
US6214987B1 (en) 1994-09-02 2001-04-10 Andrew C. Hiatt Compositions for enzyme catalyzed template-independent formation of phosphodiester bonds using protected nucleotides
US5763594A (en) 1994-09-02 1998-06-09 Andrew C. Hiatt 3' protected nucleotides for enzyme catalyzed template-independent creation of phosphodiester bonds
US5808045A (en) * 1994-09-02 1998-09-15 Andrew C. Hiatt Compositions for enzyme catalyzed template-independent creation of phosphodiester bonds using protected nucleotides
US5872244A (en) 1994-09-02 1999-02-16 Andrew C. Hiatt 3' protected nucleotides for enzyme catalyzed template-independent creation of phosphodiester bonds
WO1996011937A1 (en) * 1994-10-14 1996-04-25 Stratagene Porphyrin labeling of polynucleotides
SE9500342D0 (sv) 1995-01-31 1995-01-31 Marek Kwiatkowski Novel chain terminators, the use thereof for nucleic acid sequencing and synthesis and a method of their preparation
WO1996027025A1 (en) 1995-02-27 1996-09-06 Ely Michael Rabani Device, compounds, algorithms, and methods of molecular characterization and manipulation with molecular parallelism
EP0745686A1 (en) * 1995-06-01 1996-12-04 Roche Diagnostics GmbH The use of DNA polymerase 3'-intrinsic editing activity
EP0992511B1 (en) 1996-01-23 2009-03-11 Operon Biotechnologies, Inc. Methods and compositions for determining the sequence of nucleic acid molecules
US6312893B1 (en) * 1996-01-23 2001-11-06 Qiagen Genomics, Inc. Methods and compositions for determining the sequence of nucleic acid molecules
US6613508B1 (en) * 1996-01-23 2003-09-02 Qiagen Genomics, Inc. Methods and compositions for analyzing nucleic acid molecules utilizing sizing techniques
US6005125A (en) 1996-05-06 1999-12-21 Rutgers The State University Of New Jersey Universal allyl linker for solid-phase nucleic acid synthesis
US5821356A (en) * 1996-08-12 1998-10-13 The Perkin Elmer Corporation Propargylethoxyamino nucleotides
US5885775A (en) * 1996-10-04 1999-03-23 Perseptive Biosystems, Inc. Methods for determining sequences information in polynucleotides using mass spectrometry
CA2277545A1 (en) 1997-01-08 1998-07-16 Proligo Llc Bioconjugation of oligonucleotides
US6046005A (en) 1997-01-15 2000-04-04 Incyte Pharmaceuticals, Inc. Nucleic acid sequencing with solid phase capturable terminators comprising a cleavable linking group
WO1998033939A1 (fr) * 1997-01-31 1998-08-06 Hitachi, Ltd. Procede pour determiner une sequence de base d'acide nucleique et appareil correspondant
DE69808661T3 (de) 1997-07-28 2012-05-03 Gen-Probe Inc. Sequenzanalyse von nukleinsäuren
US6008379A (en) * 1997-10-01 1999-12-28 The Perkin-Elmer Corporation Aromatic-substituted xanthene dyes
US6485944B1 (en) 1997-10-10 2002-11-26 President And Fellows Of Harvard College Replica amplification of nucleic acid arrays
GB9815163D0 (en) 1998-07-13 1998-09-09 Brax Genomics Ltd Compounds
EP1082458A1 (en) * 1998-05-01 2001-03-14 Arizona Board Of Regents Method of determining the nucleotide sequence of oligonucleotides and dna molecules
US6780591B2 (en) * 1998-05-01 2004-08-24 Arizona Board Of Regents Method of determining the nucleotide sequence of oligonucleotides and DNA molecules
US6096875A (en) 1998-05-29 2000-08-01 The Perlein-Elmer Corporation Nucleotide compounds including a rigid linker
US6218530B1 (en) * 1998-06-02 2001-04-17 Ambergen Inc. Compounds and methods for detecting biomolecules
US6287821B1 (en) * 1998-06-11 2001-09-11 Orchid Biosciences, Inc. Nucleotide analogues with 3'-pro-fluorescent fluorophores in nucleic acid sequence analysis
US6218118B1 (en) * 1998-07-09 2001-04-17 Agilent Technologies, Inc. Method and mixture reagents for analyzing the nucleotide sequence of nucleic acids by mass spectrometry
CA2339121A1 (en) 1998-07-30 2000-02-10 Shankar Balasubramanian Arrayed biomolecules and their use in sequencing
GB0002310D0 (en) 2000-02-01 2000-03-22 Solexa Ltd Polynucleotide sequencing
US6787308B2 (en) * 1998-07-30 2004-09-07 Solexa Ltd. Arrayed biomolecules and their use in sequencing
DE19844931C1 (de) 1998-09-30 2000-06-15 Stefan Seeger Verfahren zur DNS- oder RNS-Sequenzierung
US6451525B1 (en) 1998-12-03 2002-09-17 Pe Corporation (Ny) Parallel sequencing method
US6380378B1 (en) * 1998-12-24 2002-04-30 Toagosei Company, Ltd. Nucleotide compound, nucleotide block oligonucleotide, and method for producing them
EP1163052B1 (en) 1999-02-23 2010-06-02 Caliper Life Sciences, Inc. Manipulation of microparticles in microfluidic systems
DK1159453T3 (da) * 1999-03-10 2008-10-06 Asm Scient Inc Fremgangsmåde til direkte nukleinsyresekventering
US7037654B2 (en) * 1999-04-30 2006-05-02 Aclara Biosciences, Inc. Methods and compositions for enhancing detection in determinations employing cleavable electrophoretic tag reagents
US7056661B2 (en) 1999-05-19 2006-06-06 Cornell Research Foundation, Inc. Method for sequencing nucleic acid molecules
US6818395B1 (en) * 1999-06-28 2004-11-16 California Institute Of Technology Methods and apparatus for analyzing polynucleotide sequences
US6242193B1 (en) * 1999-07-30 2001-06-05 Hitachi, Ltd. Apparatus for determining base sequence of nucleic acid
US6982146B1 (en) * 1999-08-30 2006-01-03 The United States Of America As Represented By The Department Of Health And Human Services High speed parallel molecular nucleic acid sequencing
WO2001016375A2 (en) 1999-08-30 2001-03-08 The Government Of The United States Of America, As Represented By The Secretary, Department Of Health And Human Services High speed parallel molecular nucleic acid sequencing
WO2001023610A2 (en) 1999-09-29 2001-04-05 Solexa Ltd. Polynucleotide sequencing
US6309836B1 (en) * 1999-10-05 2001-10-30 Marek Kwiatkowski Compounds for protecting hydroxyls and methods for their use
WO2001032930A1 (en) 1999-11-04 2001-05-10 California Institute Of Technology Methods and apparatuses for analyzing polynucleotide sequences
WO2001048184A2 (de) * 1999-12-23 2001-07-05 Axaron Bioscience Ag Verfahren zur parallelen sequenzierung eines nukleinsäuregemisches an einer oberfläche
GB0002389D0 (en) 2000-02-02 2000-03-22 Solexa Ltd Molecular arrays
EP1182267B1 (en) 2000-03-30 2012-01-18 Toyota Jidosha Kabushiki Kaisha Method of determining base sequence of single nucleic acid molecule
GB0013276D0 (en) * 2000-06-01 2000-07-26 Amersham Pharm Biotech Uk Ltd Nucleotide analogues
GB0016473D0 (en) 2000-07-05 2000-08-23 Amersham Pharm Biotech Uk Ltd Sequencing method
WO2002022883A1 (en) 2000-09-11 2002-03-21 The Trustees Of Columbia University In The City Of New York Combinatorial fluorescence energy transfer tags and uses thereof
ATE356222T1 (de) 2000-10-06 2007-03-15 Univ Columbia Massives parallelverfahren zur dekodierung von dna und rna
JP2004523243A (ja) 2001-03-12 2004-08-05 カリフォルニア インスティチュート オブ テクノロジー 非同期性塩基伸長によってポリヌクレオチド配列を分析するための方法および装置
US20030027140A1 (en) 2001-03-30 2003-02-06 Jingyue Ju High-fidelity DNA sequencing using solid phase capturable dideoxynucleotides and mass spectrometry
DE10120798B4 (de) 2001-04-27 2005-12-29 Genovoxx Gmbh Verfahren zur Bestimmung der Genexpression
US6613523B2 (en) * 2001-06-29 2003-09-02 Agilent Technologies, Inc. Method of DNA sequencing using cleavable tags
US7057031B2 (en) * 2001-07-13 2006-06-06 Ambergen, Inc. Nucleotide compositions comprising photocleavable markers and methods of preparation thereof
US7057026B2 (en) 2001-12-04 2006-06-06 Solexa Limited Labelled nucleotides
EP1497304B1 (en) * 2002-04-12 2014-06-25 Catalyst Assets LLC Dual-labeled nucleotides
US7074597B2 (en) * 2002-07-12 2006-07-11 The Trustees Of Columbia University In The City Of New York Multiplex genotyping using solid phase capturable dideoxynucleotides and mass spectrometry
US7414116B2 (en) * 2002-08-23 2008-08-19 Illumina Cambridge Limited Labelled nucleotides
SI3587433T1 (sl) * 2002-08-23 2020-08-31 Illumina Cambridge Limited Modificirani nukleotidi
GB0321306D0 (en) 2003-09-11 2003-10-15 Solexa Ltd Modified polymerases for improved incorporation of nucleotide analogues
US7067031B2 (en) * 2003-12-03 2006-06-27 Dew Engineering And Development Limited Process for making a ceramic armor plate
ES2639938T5 (es) 2011-09-23 2021-05-07 Illumina Inc Métodos y composiciones para la secuenciación de ácidos nucleicos

Cited By (24)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101631796A (zh) * 2007-03-09 2010-01-20 独立行政法人理化学研究所 具有由单核苷或单核苷酸衍生的结构的化合物、核酸、标记物以及核酸检测方法和试剂盒
US9410196B2 (en) 2008-09-03 2016-08-09 Quantumdx Group Limited Methods and kits for nucleic acid sequencing
US10759824B2 (en) 2008-09-03 2020-09-01 Quantumdx Group Limited Design, synthesis and use of synthetic nucleotides comprising charge mass tags
US8871921B2 (en) 2008-09-03 2014-10-28 Quantumdx Group Ltd. Design, synthesis and use of synthetic nucleotides comprising charge mass tags
US9938573B2 (en) 2008-09-03 2018-04-10 Quantumdx Group Limited Methods and kits for nucleic acid sequencing
CN103087131A (zh) * 2013-01-15 2013-05-08 上海交通大学 可逆终端及其合成和在dna合成测序中的用途
CN103087131B (zh) * 2013-01-15 2015-07-08 上海交通大学 可逆终端及其合成和在dna合成测序中的用途
CN108864232B (zh) * 2013-03-15 2022-01-04 伊鲁米纳剑桥有限公司 修饰的核苷或核苷酸
CN105377869A (zh) * 2013-03-15 2016-03-02 伊鲁米纳剑桥有限公司 修饰的核苷或核苷酸
CN108864232A (zh) * 2013-03-15 2018-11-23 伊鲁米纳剑桥有限公司 修饰的核苷或核苷酸
CN103484106B (zh) * 2013-09-05 2015-08-19 上海交通大学 四色荧光标记可逆终端及其在dna测序中的用途
CN103484106A (zh) * 2013-09-05 2014-01-01 上海交通大学 四色荧光标记可逆终端及其在dna测序中的用途
CN106117288A (zh) * 2015-05-08 2016-11-16 桑特里莱恩科技控股公司 二硫键连接的可逆终止子
US11180522B2 (en) 2015-05-08 2021-11-23 Centrillion Technology Holdings Corporation Disulfide-linked reversible terminators
CN106117288B (zh) * 2015-05-08 2019-10-15 生捷科技控股公司 二硫键连接的可逆终止子
CN114989235A (zh) * 2015-09-28 2022-09-02 哥伦比亚大学董事会 用作dna合成测序的可逆终止物的基于新的二硫键接头的核苷酸的设计与合成
US11180523B2 (en) 2016-01-04 2021-11-23 Quantumdx Group Limited Design, synthesis and use of synthetic nucleotides comprising charge mass tags
CN109562142A (zh) * 2016-04-04 2019-04-02 纽约哥伦比亚大学董事会 使用核苷酸类似物和拉曼检测的dna合成测序
WO2019042016A1 (zh) * 2017-08-31 2019-03-07 深圳华大智造科技有限公司 一种核酸探针以及一种核酸测序方法
CN108822173B (zh) * 2018-06-26 2021-04-02 合肥华纳生物医药科技有限公司 一种荧光标记可剪切核苷酸、合成方法及其在dna测序中的用途
CN108822173A (zh) * 2018-06-26 2018-11-16 合肥华纳生物医药科技有限公司 一种荧光标记可剪切核苷酸、合成方法及其在dna测序中的用途
WO2021031109A1 (zh) * 2019-08-20 2021-02-25 深圳华大智造极创科技有限公司 一种基于发光标记物光信号动力学及二次发光信号对多核苷酸进行测序的方法
CN114286867A (zh) * 2019-08-20 2022-04-05 深圳华大智造极创科技有限公司 一种基于发光标记物光信号动力学及二次发光信号对多核苷酸进行测序的方法
CN114286867B (zh) * 2019-08-20 2024-04-16 青岛华大智造科技有限责任公司 一种基于发光标记物光信号动力学及二次发光信号对多核苷酸进行测序的方法

Also Published As

Publication number Publication date
US7566537B2 (en) 2009-07-28
WO2003048387A3 (en) 2003-10-16
EP2338893A1 (en) 2011-06-29
EP1451351B1 (en) 2017-02-01
US7427673B2 (en) 2008-09-23
EP3858845A1 (en) 2021-08-04
ES2620131T3 (es) 2017-06-27
EP3266791A1 (en) 2018-01-10
HK1248710A1 (zh) 2018-10-19
EP3266791B1 (en) 2021-01-27
IS7291A (is) 2004-06-01
KR20050044668A (ko) 2005-05-12
IL162170A0 (en) 2005-11-20
DK3266791T3 (da) 2021-03-01
DK1451351T3 (en) 2017-03-27
JP2005511058A (ja) 2005-04-28
CY1123882T1 (el) 2022-05-27
EP1451351A2 (en) 2004-09-01
EP2338893B1 (en) 2017-06-28
US20060188901A1 (en) 2006-08-24
BR0214683A (pt) 2004-11-23
US20100028885A1 (en) 2010-02-04
PT3266791T (pt) 2021-03-03
PT1451351T (pt) 2017-03-29
SI3266791T1 (sl) 2021-04-30
WO2003048387A2 (en) 2003-06-12
US7785796B2 (en) 2010-08-31
US7057026B2 (en) 2006-06-06
US20030104437A1 (en) 2003-06-05
ES2858359T3 (es) 2021-09-30
AU2002349155A1 (en) 2003-06-17
CY1118765T1 (el) 2017-07-12
CA2469066A1 (en) 2003-06-12
EP2338893B9 (en) 2019-12-25
US20060160075A1 (en) 2006-07-20

Similar Documents

Publication Publication Date Title
CN1617937A (zh) 标记过的核苷酸
US10519496B2 (en) Labelled nucleotides
US7414116B2 (en) Labelled nucleotides

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C02 Deemed withdrawal of patent application after publication (patent law 2001)
WD01 Invention patent application deemed withdrawn after publication