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21. 发明申请

WO2018136497A1 LOW-NOISE BIOMOLECULAR SENSORS 审中-公开
公开(公告)号：WO2018136497A1
公开(公告)日：2018-07-26
申请号：PCT/US2018/014025
申请日：2018-01-17
申请人： ROCHE SEQUENCING SOLUTIONS, INC. , F. HOFFMAN-LA ROCHE AG , ROCHE DIAGNOSTICS GMBH
发明人： WANG, Chao
IPC分类号： G01N27/414 , G01N33/487
摘要： A method for forming a nanopore device includes providing a sapphire substrate and forming oxide layers on the front and back sides of the sapphire substrate. The oxide layer on the back is patterned to form an etch mask. The method also includes performing a crystalline orientation dependent wet anisotropic etch on the backside of the sapphire substrate using the etch mask to form a cavity having sloped sides to expose a portion of the first oxide layer. A silicon nitride membrane layer is formed on the oxide layer on the front side of the sapphire substrate. Next, the exposed portion of the oxide layer in the cavity is removed to cause the exposed portion of the silicon nitride membrane layer to be suspended over the cavity in the sapphire substrate. Subsequently, an opening is formed in the suspended portion of the silicon nitride membrane layer to form the nanopore.

22. 发明申请

WO2018108328A1 METHOD FOR INCREASING THROUGHPUT OF SINGLE MOLECULE SEQUENCING BY CONCATENATING SHORT DNA FRAGMENTS 审中-公开
公开(公告)号：WO2018108328A1
公开(公告)日：2018-06-21
申请号：PCT/EP2017/057975
申请日：2017-04-04
申请人： F. HOFFMANN-LA ROCHE AG , ROCHE DIAGNOSTICS GMBH , ROCHE SEQUENCING SOLUTIONS, INC.
发明人： SCHLECHT, Ulrich , MOK, Janine , SO, Austin
IPC分类号： C12Q1/68
摘要： The invention comprises a novel method and compositions for sequencing library preparation, which increases the throughput of single-molecule sequencing (SMS) platforms by generating long concatenated templates from pools of short DNA molecules.

23. 发明申请

WO2018091419A1 COMPOSITIONS AND METHODS FOR PROGNOSING AND TREATING COLORECTAL CANCER 审中-公开
公开(公告)号：WO2018091419A1
公开(公告)日：2018-05-24
申请号：PCT/EP2017/079084
申请日：2017-11-13
申请人： VENTANA MEDICAL SYSTEMS, INC. , F. HOFFMANN-LA ROCHE AG , ROCHE SEQUENCING SOLUTIONS INC. , ROCHE DIAGNOSTICS GMBH , CHARITÉ - UNIVERSITÄTSMEDIZIN BERLIN GLIEDKÖRPERSCHAFT DER FREIEN UNIVERSITÄT BERLIN UND DER HUMBOLDT-UNIVERSITÄT ZU BERLIN
发明人： HERRMANN, Pia , ILM, Katharina , LEITH, Katherine F. , MURANYI, Andrea , ROHR, Ulrich-Peter , SHANMUGAM, Kandavel , SINGH, Shalini , STEIN, Ulrike
IPC分类号： C12Q1/6886 , G01N33/574
摘要： A combination of mismatch repair (MMR) and Metastasis Associated in Colon Cancer 1 (MACC1) gene expression status of the patient serve as a basis for risk stratification of early stage colon cancer patients. Patients with defective MMR (dMMR) status have improved survival and do not benefit from 5-fluorouracil (5-FU) therapies. In contrast, patients with a proficient MMR (pMMR) status have a higher risk of recurrence and worse survival. The pMMR patients are then further stratified on the basis of MACC1 gene expression. Patients with a pMMR status and a low MACC1 expression have a favorable prognosis similar to patients having a dMMR status, whereas patients having a pMMR status and high MACC1 expression have a less favorable prognosis.

24. 发明申请

WO2018069484A2 MOLECULAR DETECTION AND COUNTING USING NANOPORES 审中-公开
标题翻译：用纳米孔分子检测和计数
公开(公告)号：WO2018069484A2
公开(公告)日：2018-04-19
申请号：PCT/EP2017/076152
申请日：2017-10-13
申请人： F. HOFFMANN-LA ROCHE AG , ROCHE DIAGNOSTICS GMBH , ROCHE SEQUENCING SOLUTIONS, INC.
发明人： DAVIS, Randall
IPC分类号： C12Q1/6851
CPC分类号： G01N33/48721 , C12Q1/6806 , C12Q1/6813 , C12Q1/6844 , C12Q1/6851 , C12Q1/686 , C12Q2600/16 , C12Q2533/101 , C12Q2563/179 , C12Q2565/631
摘要： Provided herein are methods and compositions for detecting and/or quantitating target analytes, including nucleic acids and polypeptides, using nanopore detectable barcodes.
摘要翻译：本文提供了使用纳米孔可检测条形码来检测和/或定量目标分析物（包括核酸和多肽）的方法和组合物。

25. 发明申请

WO2018054970A2 POL6 POLYMERASE VARIANTS 审中-公开
标题翻译： POL6聚合酶变体
公开(公告)号：WO2018054970A2
公开(公告)日：2018-03-29
申请号：PCT/EP2017/073767
申请日：2017-09-20
申请人： F. HOFFMANN-LA ROCHE AG , ROCHE DIAGNOSTICS GMBH , ROCHE SEQUENCING SOLUTIONS, INC. , ROCHE MOLECULAR SYSTEMS, INC.
发明人： BOENITZ-DULAT, Mara , SUKO, Shawn , AYER, Aruna , WOERSDOERFER, Bigna , ECKERT, Barbara , SARVABHOWMAN, Preethi , SCHWAB, Charles Wayan , WUNDERLICH, David , CLEOMA, Arnold , LEDERMAN, Ilya , THAI, Eileen , MCGAW, Colin , AMATO, Evan
IPC分类号： C12N9/12
摘要： The present disclosure provides variant Pol6 polymerase polypeptides, compositions comprising the Pol6 variant polypeptides, and methods for using the variant Pol6 polypeptides for determining the sequencing of nucleic acids, for example, by nanopore sequencing. The variant Pol6 polymerases possess decreased rates of dissociation of template from the polymerase-template complex, which result in increased processivity relative to the parental Pol6 polypeptides from which they are derived.
摘要翻译：本公开内容提供了变体Pol6聚合酶多肽，包含Pol6变体多肽的组合物，以及使用变体Pol6多肽来确定核酸测序的方法，例如通过纳米孔测序。变体Pol6聚合酶具有降低的来自聚合酶 - 模板复合物的模板解离速率，这导致相对于它们所源自的亲本Pol6多肽而言增加的持续合成能力。

26. 发明申请

WO2018050730A1 NANOPORE-BASED SEQUENCING USING VOLTAGE MODE WITH HYBRID MODE STIMULI 审中-公开
标题翻译：使用混合模式刺激的电压模式的基于纳米孔的测序
公开(公告)号：WO2018050730A1
公开(公告)日：2018-03-22
申请号：PCT/EP2017/073095
申请日：2017-09-14
申请人： F. HOFFMANN-LA ROCHE AG , ROCHE DIAGNOSTICS GMBH , ROCHE SEQUENCING SOLUTIONS, INC.
发明人： CHEN, Roger J.A. , TIAN, Hui , MANEY, J. William Jr.
IPC分类号： C12Q1/68
摘要： A liquid voltage is applied to a first side of a lipid bilayer. The liquid voltage comprises a tag-reading period with a tag-reading voltage that tends to capture a tag into a nanopore in the lipid bilayer and an open-channel period with an open-channel voltage that tends to repel the tag. A pre-charging voltage source is connected to an integrating capacitor and a working electrode on a second side of the lipid bilayer during a pre-charging time period, such that the integrating capacitor and the working electrode are charged to a pre-charging voltage. The pre-charging voltage source is disconnected from the integrating capacitor and the working electrode during an integrating time period, such that a voltage of the integrating capacitor and a voltage of the working electrode may vary as a current flows through the nanopore. The pre-charging time period overlaps with a beginning portion of the tag-reading period.
摘要翻译：将液体电压施加到脂质双层的第一面。液体电压包括具有标签读取电压的标签读取周期，标签读取电压倾向于将标签捕获到脂质双层中的纳米孔中，以及具有倾向于排斥标签的开放通道电压的开放通道周期。预充电电压源在预充电时间段期间连接到脂质双层的第二侧上的积分电容器和工作电极，使得积分电容器和工作电极被充电到预充电电压。预充电电压源在积分时间段期间从积分电容器和工作电极断开，使得积分电容器的电压和工作电极的电压可随着电流流过纳米孔而变化。预充电时间周期与标签读取周期的开始部分重叠。

27. 发明申请

WO2018002125A1 LONG LIFETIME ALPHA-HEMOLYSIN NANOPORES 审中-公开
标题翻译：长寿命α-溶血素纳米孔
公开(公告)号：WO2018002125A1
公开(公告)日：2018-01-04
申请号：PCT/EP2017/065972
申请日：2017-06-28
申请人： F. HOFFMANN-LA ROCHE AG , ROCHE DIAGNOSTICS GMBH , ROCHE SEQUENCING SOLUTIONS, INC.
发明人： CRAIG, Timothy , HARRIS, Corissa , AMBROSO, Mark , PORTER, Marshall , DIPIETRO, Matthew
IPC分类号： C07K14/31
摘要： Described herein are variants of alpha-hemolysin having at least one amino acid substitution at H35G, E111N, M113A, and/or K147N in the mature, wild-type alpha-hemolysin amino acid sequence. In certain examples, the variant may have a substitution at E111S, M113S, T145S, K147S, or L135I in the mature alpha-hemolysin amino acid sequence. The α-hemolysin variants may also include a substitution at H144A and/or a series of glycine residues spanning residues 127 to 131 of the mature, wild-type alpha hemolysin. Also provided are nanopore assemblies including the alpha-hemolysin variants, the assembly having an increased nanopore lifetime. Further, provided are variants that, in addition to providing increased lifetime, provide a decreased time-to-thread. Hence, the variants provided herein both increase nanopore lifetime and improve efficiency and accuracy of DNA sequencing reactions using nanopores comprising the variants.
摘要翻译：本文描述了在成熟的野生型α-溶血素氨基酸序列中在H35G，E111N，M113A和/或K147N处具有至少一个氨基酸取代的α-溶血素的变体。在某些实例中，变体可在成熟α-溶血素氨基酸序列中的E111S，M113S，T145S，K147S或L135I处具有取代。 α-溶血素变体还可以包括在成熟的野生型α溶血素的H144A和/或跨越残基127至131的一系列甘氨酸残基处的取代。还提供了包括α-溶血素变体的纳米孔组件，该组件具有增加的纳米孔寿命。此外，提供了除了提供增加的使用寿命之外还提供减少的线程时间的变体。因此，本文提供的变体都使用包含变体的纳米孔增加纳米孔寿命并提高DNA测序反应的效率和准确度。

28. 发明申请

WO2017210469A2 IMMUNO-PETE 审中-公开
公开(公告)号：WO2017210469A2
公开(公告)日：2017-12-07
申请号：PCT/US2017/035516
申请日：2017-06-01
申请人： F. HOFFMAN-LA ROCHE AG , ROCHE DIAGNOSTICS GMBH , ROCHE SEQUENCING SOLUTIONS, INC.
发明人： BERKA, Jan , GODWIN, Brian , DALLETT, Carolina , OZTURK, Sedide , PLATZER, Joseph , KIM, Seoyoung
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6881 , C12N15/1065 , C12Q1/68 , C12Q1/6886 , C12Q2600/16 , C12Q2525/161 , C12Q2525/186 , C12Q2537/143
摘要： Methods and compositions are described herein for primer extension target enrichment of immune receptor (BCR or TCR) sequences.
摘要翻译：本文描述了用于引物延伸目标富集免疫受体（BCR或TCR）序列的方法和组合物。

29. 发明申请

WO2017203059A1 TAGGED MULTI-NUCLEOTIDES USEFUL FOR NUCLEIC ACID SEQUENCING 审中-公开
标题翻译：标记的多核苷酸用于核酸测序
公开(公告)号：WO2017203059A1
公开(公告)日：2017-11-30
申请号：PCT/EP2017/062886
申请日：2017-05-29
申请人： F. HOFFMANN-LA ROCHE AG , ROCHE DIAGNOSTICS GMBH , ROCHE SEQUENCING SOLUTIONS, INC.
发明人： AYER, Aruna , GREMYACHINSKIY, Dmitriy , CRISALLI, Peter , TAING, Meng
IPC分类号： C07H21/00 , C12Q1/68
摘要： The present disclosure relates to tagged multi-nucleotide compounds, which comprise a single tag moiety covalently linked to a plurality of nucleoside-5'-oligophosphate moieties. As disclosed herein, these tagged multi-nucleotide compounds have improved characteristics as polymerase substrates and can be used in a range of nucleic acid detection and sequencing methods, including nanopore sequencing-by-synthesis.
摘要翻译：本公开涉及标记的多核苷酸化合物，其包含与多个核苷-5'-寡磷酸酯部分共价连接的单个标签部分。如本文所公开的，这些标记的多核苷酸化合物作为聚合酶底物具有改善的特征，并且可以用于一系列核酸检测和测序方法，包括纳米孔合成测序。

30. 发明申请

WO2017202917A1 TAGGED MULTI-NUCLEOTIDES USEFUL FOR NUCLEIC ACID SEQUENCING 审中-公开
公开(公告)号：WO2017202917A1
公开(公告)日：2017-11-30
申请号：PCT/EP2017/062552
申请日：2017-05-24
申请人： F. HOFFMANN-LA ROCHE AG , ROCHE DIAGNOSTICS GMBH , ROCHE SEQUENCING SOLUTIONS, INC.
发明人： AYER, Aruna , GREMYACHINSKIY, Dmitriy , CRISALLI, Peter , TAING, Meng
IPC分类号： C07H21/00 , C12Q1/68
摘要： The present disclosure relates to tagged multi-nucleotide compounds, which comprise a single tag moiety covalently linked to a plurality of nucleoside-5'-oligophosphate moieties. As disclosed herein, these tagged multi-nucleotide compounds have improved characteristics as polymerase substrates and can be used in a range of nucleic acid detection and sequencing methods, including nanopore sequencing-by-synthesis.

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