公开(公告)号：US20070054285A1

公开(公告)日：2007-03-08

申请号：US11399310

申请日：2006-04-06

申请人： Kevin McKernan ,  Junaid Ziauddin

发明人： Kevin McKernan ,  Junaid Ziauddin

IPC分类号： C12Q1/68 ,  C07H21/04

CPC分类号： C12N15/1006 ,  C12N15/1013

摘要： Described herein is a method in which genomic nucleic acid of a cell can be separated from nucleic acid having a molecular weight that is lower than the molecular weight of the genomic nucleic acid (e.g., plasmid DNA) of the cell directly from a cell growth culture. Also described herein, a method in which genomic nucleic acid can be separated from nucleic acid having a molecular weight that is lower than the molecular weight of the genomic nucleic acid in a cell lysate without the need to prepare a cleared lysate.

摘要翻译： 本文描述了一种方法，其中细胞的基因组核酸可从直接来自细胞生长培养物的细胞的分子量低于细胞的基因组核酸（例如质粒DNA）的分子量的核酸分离 。 本文还描述了可以将基因组核酸与细胞裂解物中分子量低于基因组核酸分子量的核酸分离而不需要制备澄清的裂解物的方法。

公开(公告)号：US20060177836A1

公开(公告)日：2006-08-10

申请号：US11194072

申请日：2005-07-29

申请人： Kevin McKernan ,  Erik Gustafson ,  Adrianne Brand

发明人： Kevin McKernan ,  Erik Gustafson ,  Adrianne Brand

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6806 ,  C12N15/1006 ,  C12N15/1013 ,  C12Q1/6834 ,  C12Q1/6869 ,  C12Q2527/125

摘要： The present invention is directed to a method of isolating a target species (e.g., target nucleic acid species) from a mixture. In the methods of the present invention, the mixture is combined with solid phase carriers having a surface comprising multiple functional groups one of which reversibly and selectively binds the target species. In a particular embodiment, the mixture is combined with solid phase carriers having a first functional group which reversibly binds nucleic acids and a second functional group which selectively and reversibly binds the target nucleic acid species, thereby producing a first combination. The first combination is maintained under conditions appropriate for binding of the nucleic acids to the first functional group and binding of the target nucleic acid species to the second functional group. The solid phase carriers are separated from the first combination, and combined with an agent (e.g., buffer) that selectively removes (e.g., elutes) either the nucleic acid from the first functional group or the target nucleic acid species from the second functional group of the solid phase carriers, thereby isolating a target nucleic acid species from a mixture comprising a plurality of nucleic acid species.

公开(公告)号：US20060003357A1

公开(公告)日：2006-01-05

申请号：US11129218

申请日：2005-05-13

申请人： Kevin McKernan ,  Paul McEwan ,  William Morris

发明人： Kevin McKernan ,  Paul McEwan ,  William Morris

IPC分类号： C12Q1/68 ,  C12N1/08

CPC分类号： C12N15/1013 ,  C12N15/1006 ,  C12Q1/6806 ,  C12Q2563/143 ,  C12Q2527/137 ,  C12Q2523/308

摘要： A method of isolating target nucleic acid molecules from a solution comprising a mixture of different size nucleic acid molecules, in the presence or absence of other biomolecules, by selectively facilitating the adsorption of a particular species of nucleic acid molecule to the functional group-coated surface of magnetically responsive paramagnetic microparticles is disclosed. Separation is accomplished by manipulating the ionic strength and polyalkylene glycol concentration of the solution to selectively precipitate, and reversibly adsorb, the target species of nucleic acid molecule, characterized by a particular molecular size, to paramagnetic microparticles, the surfaces of which act as a bioaffinity adsorbent for the nucleic acids. The target nucleic acid is isolated from the starting mixture based on molecular size and through the removal of magnetic beads to which the target nucleic acid molecules have been adsorbed. The disclosed method provides a simple, robust and readily automatable means of nucleic acid isolation and purification which produces high quality nucleic acid molecules suitable for: capillary electrophoresis, nucleotide sequencing, reverse transcription cloning the transfection, transduction or microinjection of mammalian cells, gene therapy protocols, the in vitro synthesis of RNA probes, cDNA library construction and PCR amplification.

公开(公告)号：US20060078923A1

公开(公告)日：2006-04-13

申请号：US11231363

申请日：2005-09-20

申请人： Kevin McKernan ,  Junaid Ziauddin

发明人： Kevin McKernan ,  Junaid Ziauddin

IPC分类号： C12Q1/68 ,  C12N1/08

CPC分类号： C12N15/1006 ,  C12N15/1013

摘要： Described herein is a method in which genomic nucleic acid of a cell can be separated from nucleic acid having a molecular weight that is lower than the molecular weight of the genomic nucleic acid (e.g., plasmid DNA) of the cell directly from a cell growth culture. Also described herein, a method in which genomic nucleic acid can be separated from nucleic acid having a molecular weight that is lower than the molecular weight of the genomic nucleic acid in a cell lysate without the need to prepare a cleared lysate. The present invention is also directed to a method of isolating genomic nucleic acid (e.g., RNA, DNA) of a cell or an organism.

摘要翻译： 本文描述了一种方法，其中细胞的基因组核酸可从直接来自细胞生长培养物的细胞的分子量低于细胞的基因组核酸（例如质粒DNA）的分子量的核酸分离 。 本文还描述了可以将基因组核酸与细胞裂解物中分子量低于基因组核酸分子量的核酸分离而不需要制备澄清的裂解物的方法。 本发明还涉及分离细胞或生物体的基因组核酸（例如RNA，DNA）的方法。

公开(公告)号：US20100285461A1

公开(公告)日：2010-11-11

申请号：US12632713

申请日：2009-12-07

申请人： Douglas R. Smith ,  Kevin McKernan

发明人： Douglas R. Smith ,  Kevin McKernan

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6869 ,  C12Q2525/113

摘要： The present invention encompasses methods for producing a modified polynucleotide sequence that comprises a (e.g., one or more) phosphorothiolate linkage, methods for determining a polynucleotide sequence comprising a (e.g., one or more) phosphorothiolate linkage, and methods for separating forward and reverse extension products that comprise a (e.g., one or more) phosphorothiolate linkage. The invention also encompasses kits for producing and/or determining the sequence of a modified polynucleotide that comprises a (e.g., one or more) phosphorothiolate linkage.

摘要翻译： 本发明包括用于产生包含（例如一个或多个）硫代硫醇键的修饰的多核苷酸序列的方法，用于测定包含（例如一个或多个）硫代硫醇键的多核苷酸序列的方法，以及用于分离正向和反向延伸的方法 包括（例如，一个或多个）硫代硫醇键的产物。 本发明还包括用于产生和/或确定包含（例如一个或多个）硫代磷酸酯键的修饰多核苷酸序列的试剂盒。

公开(公告)号：US20140243232A1

公开(公告)日：2014-08-28

申请号：US14232572

申请日：2012-07-13

申请人： Gavin Meredith ,  Christopher Clouser ,  Tanya Sokolsky ,  Kimberly Mather ,  Kevin McKernan ,  Marie Callahan ,  Gary Bee

发明人： Gavin Meredith ,  Christopher Clouser ,  Tanya Sokolsky ,  Kimberly Mather ,  Kevin McKernan ,  Marie Callahan ,  Gary Bee

IPC分类号： C12Q1/68 ,  C07H21/00

CPC分类号： C12Q1/6874 ,  C12Q1/6832 ,  C12Q2525/191 ,  C12Q2537/159 ,  C12Q2537/163 ,  C12Q2563/131

摘要： In some embodiments, the present teachings provide compositions, systems, methods and kits for reducing the complexity of nucleotide sequences in a nucleic acid sample comprising the steps: hybridizing a plurality of polynucleotide constructs to at least one blocker oligonucleotide and to at least one capture oligonucleotide, wherein the plurality of polynucleotide constructs include a plurality of polynucleotides each joined to at least one nucleic acid adaptor, wherein the at least one nucleic acid adaptor can hybridize to the at least one blocker oligonucleotide, and wherein the at least one capture oligonucleotide can hybridize to at least a portion of target polynucleotides that are a sub-population of the plurality of polynucleotides, so as to produce a capture duplex.

摘要翻译： 在一些实施方案中，本教导提供了用于降低核酸样品中核苷酸序列的复杂性的组合物，系统，方法和试剂盒，其包括以下步骤：将多个多核苷酸构建体与至少一个阻断寡核苷酸和至少一个捕获寡核苷酸杂交 其中所述多个多核苷酸构建体包括多个多核苷酸，其各自与至少一个核酸衔接子连接，其中所述至少一个核酸衔接子可以与所述至少一个阻断寡核苷酸杂交，并且其中所述至少一个捕获寡核苷酸可以杂交 至少一部分作为多个多核苷酸的亚群的靶多核苷酸，以产生捕获双链体。

公开(公告)号：US08058030B2

公开(公告)日：2011-11-15

申请号：US12632713

申请日：2009-12-07

申请人： Douglas R. Smith ,  Kevin McKernan

发明人： Douglas R. Smith ,  Kevin McKernan

IPC分类号： C12P19/34 ,  C12Q1/68 ,  C07H21/02 ,  C07H21/04

CPC分类号： C12Q1/6869 ,  C12Q2525/113

摘要： The present invention encompasses methods for producing a modified polynucleotide sequence that comprises a (e.g., one or more) phosphorothiolate linkage, methods for determining a polynucleotide sequence comprising a (e.g., one or more) phosphorothiolate linkage, and methods for separating forward and reverse extension products that comprise a (e.g., one or more) phosphorothiolate linkage. The invention also encompasses kits for producing and/or determining the sequence of a modified polynucleotide that comprises a (e.g., one or more) phosphorothiolate linkage.

摘要翻译： 本发明包括用于产生包含（例如一个或多个）硫代硫醇键的修饰的多核苷酸序列的方法，用于测定包含（例如一个或多个）硫代硫醇键的多核苷酸序列的方法，以及用于分离正向和反向延伸的方法 包括（例如，一个或多个）硫代硫醇键的产物。 本发明还包括用于产生和/或确定包含（例如一个或多个）硫代磷酸酯键的修饰多核苷酸序列的试剂盒。

公开(公告)号：US20100120034A1

公开(公告)日：2010-05-13

申请号：US12498285

申请日：2009-07-06

申请人： Kevin McKernan ,  Benjamin Schroeder ,  Victoria Boyd

发明人： Kevin McKernan ,  Benjamin Schroeder ,  Victoria Boyd

IPC分类号： C12Q1/68 ,  C07H21/04

CPC分类号： C12Q1/6827 ,  C12Q1/686 ,  C12Q2525/307 ,  C12Q2525/191 ,  C12Q2521/331

摘要： Various embodiments of the present teachings relate to methods for the methylation analysis of nucleic acids. The subject methods include methods that result in the preparation of mate-pair libraries suitable for highly multiplexed DNA sequencing. Embodiments include methods of preparing mate-pair libraries comprising a first tag sequence and a second tag sequence, wherein one of the tag sequences has been converted by a methylation conversion agent and the other tag sequence has not been converted by the methylation conversion agent. Other embodiments provided include intermediates for making the mate-pair library and kits for making the mate-pair libraries. Also provided is software and computer systems for analyzing the methylation levels of genomic DNA from which the tag sequences were derived.

摘要翻译： 本发明的各种实施方案涉及核酸甲基化分析的方法。 主题方法包括导致适合于高度多重化DNA测序的配偶对文库的制备的方法。 实施方案包括制备包含第一标签序列和第二标签序列的伴侣对文库的方法，其中标签序列之一已被甲基化转化剂转化，另一个标签序列尚未被甲基化转化剂转化。 提供的其它实施方案包括用于制备配对文库和用于制备配对文库的试剂盒的中间体。 还提供了用于分析衍生标签序列的基因组DNA的甲基化水平的软件和计算机系统。

公开(公告)号：US06534262B1

公开(公告)日：2003-03-18

申请号：US09311317

申请日：1999-05-13

申请人： Kevin McKernan ,  Paul McEwan ,  William Morris

发明人： Kevin McKernan ,  Paul McEwan ,  William Morris

IPC分类号： C12Q168

CPC分类号： C12N15/1013 ,  C12N15/1006 ,  C12Q1/6806 ,  C12Q2563/143 ,  C12Q2527/137 ,  C12Q2523/308

摘要： A method of isolating target nucleic acid molecules from a solution comprising a mixture of different size nucleic acid molecules, in the presence or absence of other biomolecules, by selectively facilitating the adsorption of a particular species of nucleic acid molecule to the functional group-coated surface of magnetically responsive paramagnetic microparticles is disclosed. Separation is accomplished by manipulating the ionic strength and polyalkylene glycol concentration of the solution to selectively precipitate, and reversibly adsorb, the target species of nucleic acid molecule, characterized by a particular molecular size, to paramagnetic microparticles, the surfaces of which act as a bioaffinity adsorbent for the nucleic acids. The target nucleic acid is isolated from the starting mixture based on molecular size and through the removal of magnetic beads to which the target nucleic acid molecules have been adsorbed. The disclosed method provides a simple, robust and readily automatable means of nucleic acid isolation and purification which produces high quality nucleic acid molecules suitable for: capillary electrophoresis, nucleotide sequencing, reverse transcription cloning the transfection, transduction or microinjection of mammalian cells, gene therapy protocols, the in vitro synthesis of RNA probes, cDNA library construction and PCR amplification.

摘要翻译： 在存在或不存在其它生物分子的情况下，通过选择性促进特定种类的核酸分子吸附到官能团涂覆的表面上，从包含不同大小的核酸分子的混合物的溶液中分离靶核酸分子的方法 的磁响应顺磁性微粒。 通过操作溶液的离子强度和聚亚烷基二醇浓度来选择性地沉淀并可逆吸附具有特定分子尺寸的特征的核酸分子的靶物种到顺磁性微粒，其表面作为生物亲和力 用于核酸的吸附剂。 基于分子大小和去除已经吸附了靶核酸分子的磁珠，从起始混合物中分离靶核酸。 所公开的方法提供了简单，稳健且易于自动化的核酸分离和纯化手段，其产生适合于毛细管电泳，核苷酸测序，逆转录克隆转染，转导或显微注射哺乳动物细胞的高质量核酸分子，基因治疗方案 ，RNA探针的体外合成，cDNA文库构建和PCR扩增。

公开(公告)号：US20140057251A1

公开(公告)日：2014-02-27

申请号：US13588935

申请日：2012-08-17

申请人： Kevin McKernan

发明人： Kevin McKernan

IPC分类号： C07K16/40 ,  C12Q1/68 ,  G01N33/573 ,  C12N9/88

CPC分类号： C07K16/40 ,  C12N9/88 ,  C12Q1/68 ,  C12Q1/6895 ,  C12Q2600/13 ,  C12Q2600/156 ,  C12Q2600/158 ,  G01N33/573

摘要： Using the efficiency of next generation sequencing, a draft de novo reference sequence for the Cannabis (C.) Sativa and C. Indica genomes has been generated as well as four full length contiguous sequences with homology to THCA and CBDA synthases and 10 partially homologous contigs with truncated ORFs. In particular aspects the invention is directed to an (one or more) isolated sequence (e.g., nucleic acid sequence, DNA, RNA, genomic sequence, polypeptide) of a Cannabis genome and uses thereof.

摘要翻译： 使用下一代测序的效率，已经产生了大麻（C.）Sativa和C.Inica基因组的从头参考序列草案，以及与THCA和CBDA合酶和10个部分同源的重叠群具有同源性的四个全长连续序列 与截断的ORF。 在具体方面，本发明涉及大麻基因组的（一个或多个）分离的序列（例如，核酸序列，DNA，RNA，基因组序列，多肽）及其用途。