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    Sequential sequencing
    2.
    发明授权
    Sequential sequencing 有权

    公开(公告)号：US10760123B2

    公开(公告)日：2020-09-01

    申请号：US14990339

    申请日：2016-01-07

    申请人： NuGEN Technologies, Inc.

    发明人： Doug Amorese ,  Benjamin G. Schroeder ,  Jonathan Scolnick

    IPC分类号： C12Q1/6874 ,  G16B30/00

    摘要： The present invention provides improved methods, compositions and kits for short read next generation sequencing (NGS). The methods, compositions and kits of the present invention enable phasing of two or more nucleic acid sequences in a sample, i.e. determining whether the nucleic acid sequences (typically comprising regions of sequence variation) are located on the same chromosome and/or the same chromosomal fragment. Phasing information is obtained by performing multiple, successive sequencing reactions from the same immobilized nucleic acid template. The methods, compositions and kits provided herein are useful, for example, for haplotyping, SNP phasing, or for determining downstream exons in RNA-seq.

    Compositions for amplification of RNA sequences using composite primers
    4.
    发明授权
    Compositions for amplification of RNA sequences using composite primers 有权
    使用复合引物扩增RNA序列的组合物

    公开(公告)号：US09181582B2

    公开(公告)日：2015-11-10

    申请号：US13922146

    申请日：2013-06-19

    申请人： NUGEN TECHNOLOGIES, INC.

    发明人： Nurith Kurn

    IPC分类号： C07H21/04 ,  C12Q1/68

    CPC分类号： C12Q1/6853 ,  C12Q2531/119

    摘要： The invention provides methods for isothermal amplification of RNA. The methods are particularly suitable for amplifying a plurality of RNA species in a sample. The methods employ a composite primer, a second primer and strand displacement to generate multiple copies of DNA products comprising sequences complementary to an RNA sequence of interest. In another aspect, the methods employ a single primer (which is a composite primer) and strand displacement to generate multiple copies of DNA products comprising sequences complementary to an RNA sequence of interest. In some embodiments, a transcription step is included to generate multiple copies of sense RNA of an RNA sequence of interest. The methods are useful for preparation of nucleic acid libraries and substrates for analysis of gene expression of cells in biological samples. The invention also provides compositions and kits for practicing the amplification methods, as well as methods which use the amplification products.

    摘要翻译： 本发明提供了RNA等温扩增的方法。 该方法特别适用于扩增样品中的多种RNA物种。 该方法采用复合引物，第二引物和链置换，以产生包含与感兴趣的RNA序列互补的序列的DNA产物的多个拷贝。 另一方面，该方法采用单一引物（其是复合引物）和链置换以产生包含与感兴趣的RNA序列互补的序列的DNA产物的多个拷贝。 在一些实施方案中，包括转录步骤以产生感兴趣的RNA序列的有义RNA的多个拷贝。 该方法可用于制备用于分析生物样品中细胞基因表达的核酸文库和底物。 本发明还提供了用于实施扩增方法的组合物和试剂盒，以及使用扩增产物的方法。

    COMPOSITIONS AND METHODS FOR NEGATIVE SELECTION OF NON-DESIRED NUCLEIC ACID SEQUENCES
    5.
    发明申请
    COMPOSITIONS AND METHODS FOR NEGATIVE SELECTION OF NON-DESIRED NUCLEIC ACID SEQUENCES 有权
    阴性选择非靶核酸序列的组合物和方法

    公开(公告)号：US20150299767A1

    公开(公告)日：2015-10-22

    申请号：US14390012

    申请日：2013-03-15

    申请人： NUGEN TECHNOLOGIES, INC.

    发明人： Christopher Armour ,  Doug Amorese ,  Bin Li ,  Nurith Kurn

    IPC分类号： C12Q1/68

    CPC分类号： C12Q1/6806 ,  C12N15/1093 ,  C12N15/1096 ,  C12Q1/6844 ,  C12Q2525/131 ,  C12Q2525/191 ,  C12Q2521/301 ,  C12Q2521/531 ,  C12Q2537/159

    摘要： The present invention provides methods, compositions and kits for the generation of next generation sequencing (NGS) libraries in which non-desired nucleic acid sequences have been depleted or substantially reduced. The methods, compositions and kits provided herein are useful, for example, for the production of libraries from total RNA with reduced ribosomal RNA and for the reduction of common mRNA species in expression profiling from mixed samples where the mRNAs of interest are present at low levels. The methods of the invention can be employed for the elimination of non-desired nucleic acid sequences in a sequence-specific manner, and consequently, for the enrichment of nucleic acid sequences of interest in a nucleic acid library.

    摘要翻译： 本发明提供用于产生下一代测序（NGS）文库的方法，组合物和试剂盒，其中非期望的核酸序列已经耗尽或显着降低。 本文提供的方法，组合物和试剂盒是有用的，例如用于从具有降低的核糖体RNA的总RNA生产文库，并且用于从感兴趣的mRNA以低水平存在的混合样品中降低表达谱中常见的mRNA种类 。 本发明的方法可用于以序列特异性方式消除非期望的核酸序列，并因此用于富集核酸文库中目的核酸序列。

    METHODS, COMPOSITIONS AND KITS FOR GENERATION OF STRANDED RNA OR DNA LIBRARIES
    6.
    发明申请
    METHODS, COMPOSITIONS AND KITS FOR GENERATION OF STRANDED RNA OR DNA LIBRARIES 审中-公开
    方法，用于生成条带RNA或DNA文库的组合物和试剂盒

    公开(公告)号：US20140274729A1

    公开(公告)日：2014-09-18

    申请号：US14030761

    申请日：2013-09-18

    申请人： NuGEN Technologies, Inc.

    发明人： Nurith Kurn ,  Bin Li

    IPC分类号： B01J19/00

    CPC分类号： B01J19/0046

    摘要： The invention provides methods and compositions, including kits, for the construction of directional nucleic acid libraries. The invention further provides methods and compositions for the amplification and sequencing of directional cDNA libraries.

    摘要翻译： 本发明提供了用于构建定向核酸文库的方法和组合物，包括试剂盒。 本发明还提供了用于扩增和测序定向cDNA文库的方法和组合物。

    Methods for Fragmentation and Labeling of Nucleic Acids
    7.
    发明申请
    Methods for Fragmentation and Labeling of Nucleic Acids 审中-公开
    核酸分离和标记方法

    公开(公告)号：US20140065692A1

    公开(公告)日：2014-03-06

    申请号：US14012409

    申请日：2013-08-28

    申请人： NuGen Technologies, Inc.

    发明人： Nurith KURN ,  Pengchin Chen

    IPC分类号： C12Q1/68

    CPC分类号： C12Q1/6811 ,  C12Q1/6806 ,  C12Q2533/101 ,  C12Q2525/119 ,  C12Q2523/107 ,  C12Q2521/319

    摘要： The invention provides methods, compositions, and kits for fragmentation and labeling of nucleic acids. More particularly, the invention relates to methods for fragmentation of nucleic acids to produce fragments with 3′ end hydroxyl groups within a desired size range. In methods of the invention, nucleic acids are fragmented at abasic sites to produce fragments with blocked 3′ ends. The 3′ ends are unblocked to produce polynucleotide fragments with hydroxyl groups at their 3′ ends. Methods, kits, and compositions for carrying out fragmentation of a polynucleotide template in a single reaction mixture to yield fragments with 3′-hydroxl ends within the desired size range are disclosed.

    摘要翻译： 本发明提供了用于核酸的片段化和标记的方法，组合物和试剂盒。 更具体地说，本发明涉及用于在所需尺寸范围内产生具有3'末端羟基的片段的核酸断裂方法。 在本发明的方法中，核酸在脱碱基位点被片段化以产生具有封闭的3'末端的片段。 3'末端未封闭以在其3'末端产生具有羟基的多核苷酸片段。 公开了用于在单个反应混合物中进行多核苷酸模板的片段化以在所需尺寸范围内产生具有3'-羟基末端的片段的方法，试剂盒和组合物。

    Global Amplification Using a Randomly Primed Composite Primer
    8.
    发明申请
    Global Amplification Using a Randomly Primed Composite Primer 有权
    全面扩增使用随机引物复合引物

    公开(公告)号：US20140038236A1

    公开(公告)日：2014-02-06

    申请号：US13918636

    申请日：2013-06-14

    申请人： NuGEN Technologies, Inc.

    发明人： Nurith Kurn ,  Shenglong Wang

    IPC分类号： C12P19/34

    CPC分类号： C12P19/34 ,  C12Q1/6844 ,  C12Q1/6853 ,  C12Q2537/143 ,  C12Q2525/203 ,  C12Q2525/179

    摘要： The invention relates to the field of polynucleotide amplification. More particularly, the invention provides methods, compositions and kits for amplification of (i.e., making multiple copies of) a multiplicity of different polynucleotide template sequences using a randomly primed RNA/DNA composite primer.

    摘要翻译： 本发明涉及多核苷酸扩增领域。 更具体地，本发明提供了使用随机引物的RNA / DNA复合引物扩增多个不同多核苷酸模板序列（即制备多个拷贝）的方法，组合物和试剂盒。

    METHODS FOR LINEAR SAMPLE PROCESSING
    9.
    发明申请
    METHODS FOR LINEAR SAMPLE PROCESSING 审中-公开

    公开(公告)号：US20190291116A1

    公开(公告)日：2019-09-26

    申请号：US16355155

    申请日：2019-03-15

    申请人： NuGEN Technologies, Inc.

    发明人： Douglas A. Amorese

    IPC分类号： B01L9/00 ,  B01L7/00

    摘要： The invention provides methods for linear processing of multiple samples through a series of reactions. The methods allow parallel processing of multiple samples through a series of reactions. Systems for performing the methods are also provided.