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公开(公告)号：US06258542B1
公开(公告)日：2001-07-10
申请号：US09388917
申请日：1999-09-02
IPC分类号： C12Q168
CPC分类号： C12Q1/6834 , Y10S435/912
摘要： In order to make possible to preserve promptly and efficiently a DNA and to distribute the same without taking much labor and much time, a DNA solution is allowed to adhere to a sheet-like support having a prescribed thickness, and the DNA solution which has been allowed to adhere to the support is dried to fix the DNA onto the support.
摘要翻译： 为了能够迅速且有效地保存DNA并且分发相同的DNA而不需要花费太多的劳动和时间，可以使DNA溶液粘附到具有规定厚度的片状载体上，并且已经将DNA溶液 允许粘附到支持物上以将DNA固定在载体上。
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公开(公告)号：US06221599B1
公开(公告)日：2001-04-24
申请号：US09414531
申请日：1999-10-08
IPC分类号： C12Q168
CPC分类号： C12N15/1096 , C12Q1/6844 , C12Q2527/101 , C12Q2521/107 , C12Q2527/125
摘要： A method for preparing a cDNA from a mRNA using a reverse transcriptase wherein reverse transcription is performed at a temperature at which the mRNA does not take a secondary structure, for example, at a temperature of 45° C. or more. The method is performed, for example, using a heat-labile reverse transcriptase in the presence of a substance exhibiting chaperone function having chaperone function such as saccharides. The method is performed, for example, in the presence of metal ions necessary for activation of the reverse transcriptase and a chelating agent for the metal ions such as a deoxynucleotide triphosphate. The method is capable of reverse transcription over the full length of mRNA template even if the mRNA is a long chain mRNA and, as a result, producing a full length cDNA.
摘要翻译： 使用逆转录酶从mRNA制备cDNA的方法，其中在mRNA不采取二级结构的温度下进行逆转录，例如在45℃或更高的温度下进行。 该方法例如在具有伴侣伴侣功能的物质的存在下使用热不稳定的逆转录酶，例如糖类。 该方法例如在活化逆转录酶所需的金属离子和金属离子螯合剂如脱氧核苷酸三磷酸的存在下进行。 该方法能够在mRNA模板的全长上进行逆转录，即使mRNA是长链mRNA，因此产生全长cDNA。
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公开(公告)号：US5916775A
公开(公告)日：1999-06-29
申请号：US877874
申请日：1997-06-18
CPC分类号： C12Q1/6806 , C12N15/101 , C12N15/1017
摘要： Disclosed is a method for collecting DNA by lysing microbial cells, adsorbing released DNA on a carrier and collecting the DNA adsorbed on the carrier, which method comprises the following steps of (1) lysing the microbial cells in the presence of the carrier so that the DNA obtained by lysing cells is adsorbed onto the carrier, separating solutions used for lysing cells and adsorbing DNA from the carrier, and eluting the DNA adsorbed on the carrier with a solution for eluting DNA and collecting eluted DNA, or (2) feeding microbial cells into a column comprising the carrier provided on a membrane filter capable of retaining a solution and permeating the solution when aspirated, lysing the microbial cells in the column so that the DNA obtained by lysing cells is adsorbed onto the carrier, separating solutions used for lysing cells and adsorbing DNA in the previous step from the column by aspiration, and feeding a solution for eluting DNA into the column and aspirating to collect the DNA adsorbed on the carrier. The methods of the present invention enable collection of DNA by the chaotropic ion method employing an apparatus with simpler structure and fewer operations.
摘要翻译： 公开了通过溶解微生物细胞，将载体上释放的DNA吸附并收集载体上的DNA收集DNA的方法，该方法包括以下步骤：（1）在载体存在下裂解微生物细胞，使得 通过裂解细胞获得的DNA被吸附到载体上，分离用于裂解细胞并从载体吸附DNA的溶液，并用洗脱DNA并收集洗脱的DNA的溶液洗脱载体上的DNA，或（2）喂养微生物细胞 进入包含载体的柱，该载体提供在能够保留溶液并在吸出时渗透溶液的膜过滤器上，裂解塔中的微生物细胞，使得通过裂解细胞获得的DNA被吸附到载体上，用于裂解细胞的分离溶液 并通过抽吸从柱上吸附DNA，并将用于将DNA洗脱的溶液供入柱中并吸出以收集DNA 吸附在载体上。 本发明的方法能够利用离液力离子法采用具有更简单结构和更少操作的装置来收集DNA。
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44.发明授权Pretreatment method of biological sample, detection method of RNA, and pretreatment kit 有权生物样品预处理方法，RNA检测方法及预处理试剂盒公开(公告)号：US09518901B2
公开(公告)日：2016-12-13
申请号：US13807605
申请日：2012-06-28
申请人： Yoshihide Hayashizaki , Kengo Usui , Saori Goda , Kazuhito Nomura , Yuki Kawai
发明人： Yoshihide Hayashizaki , Kengo Usui , Saori Goda , Kazuhito Nomura , Yuki Kawai
CPC分类号： G01N1/34 , C12N15/1003 , C12Q1/6806
摘要： The present invention is to provide a pretreatment method that allows RNA to be detected promptly and simply. RNA degradation activity due to lactoferrin present in the human rhinal mucosa is inhibited, for example, by adding iron ion and carbonate ion to a biological sample that contains the human rhinal mucosa. With the pretreated biological sample, an RNA virus gene can be amplified by a reverse transcriptase. Iron ion and carbonate ion can also inhibit reverse transcriptase inhibition due to lysozyme C contained in the human rhinal mucosa. Further, it is preferable to remove the envelope of the RNA virus by adding SDS to the biological sample that contains the human rhinal mucosa.
摘要翻译： 本发明提供一种能够及时且简单地检测RNA的预处理方法。 例如，通过将铁离子和碳酸根离子加入到含有人类鼻粘膜的生物样品中，可抑制由于存在于人类鼻粘膜中的乳铁蛋白引起的RNA降解活性。 通过预处理的生物样品，RNA病毒基因可以通过逆转录酶扩增。 铁离子和碳酸根离子也可以抑制人类鼻粘膜中包含的溶菌酶C的逆转录酶抑制。 此外，优选通过向包含人类鼻粘膜的生物样品中加入SDS来除去RNA病毒的包膜。
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公开(公告)号：US20110243910A1
公开(公告)日：2011-10-06
申请号：US13058970
申请日：2009-07-07
IPC分类号： A61K38/45 , C12N9/96 , G01N33/573 , C12N5/071 , C12Q1/68 , C12M1/40 , A61P19/02 , A61P35/00 , A61P35/02 , A61P11/06 , A61P17/14 , A61P7/06 , A61P3/00 , C07K14/00
CPC分类号： C12N9/1276 , A61K38/00 , C12N9/127 , C12Y207/07048 , C12Y207/07049
摘要： The invention provides compositions comprising a TERT-RMRP or TERT-RNA complex and methods of treating subjects with genetic diseases in which gene silencing is either increased by administering the compositions of the invention or decreased by administering an inhibitor of the RNA-dependent RNA polymerase (RdRP) activity of these compositions. Moreover, the invention provides methods of screening for agonists and antagonists of RdRP activity and TERT-RMRP complex formation. Finally, the invention provides a method of identifying a RNA molecule that forms a complex with a TERT polypeptide and has RdRP activity.
摘要翻译： 本发明提供了包含TERT-RMRP或TERT-RNA复合物的组合物和治疗具有遗传疾病的受试者的方法，其中通过施用本发明的组合物或通过施用RNA依赖性RNA聚合酶的抑制剂降低基因沉默 RdRP）活性。 此外，本发明提供筛选RdRP活性和TERT-RMRP复合物形成的激动剂和拮抗剂的方法。 最后，本发明提供了鉴定与TERT多肽形成复合物并具有RdRP活性的RNA分子的方法。
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公开(公告)号：US07374882B2
公开(公告)日：2008-05-20
申请号：US10432991
申请日：2001-11-11
CPC分类号： C12N15/115 , C07H19/10 , C07H19/20 , C07H21/00 , C12N2310/33 , C12Q1/6869 , C40B30/04 , G01N33/6845 , C12Q2525/117
摘要： Aptamers are nucleic acids and similar molecules, such as peptide-nucleic acids, that specifically bind to a ligand such as a protein or peptide. The present invention provides aptamers comprising at least one base capable of base pairing and different from the standard Watson-Crick bases. The present invention also relates to a method for preparation of such aptamers and to methods for sequencing nucleic acids that comprise at least one base capable of base pairing and different from the standard Watson-Crick bases.
摘要翻译： 适体是核酸和类似分子，例如肽 - 核酸，其特异性结合配体如蛋白质或肽。 本发明提供了包含至少一个能够碱基配对并且不同于标准沃森 - 克里克碱基的碱基的适体。 本发明还涉及这种适体的制备方法以及核酸测序方法，所述方法包括至少一个能够碱基配对且不同于标准沃森 - 克里克碱基的碱基。
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公开(公告)号：US20080108804A1
公开(公告)日：2008-05-08
申请号：US11591682
申请日：2006-11-02
IPC分类号： C07H21/04
CPC分类号： C07H21/04 , C12N15/1096 , C12Q1/686 , C12Q2521/107 , C12Q2533/107
摘要： A method is disclosed for the modification of an end of RNA molecules and the use of such modified RNA molecules in cDNA synthesis for the purpose of cloning, detection, sequencing, and amplification of parts of the RNAs, the entire RNAs, or any cDNAs derived from such modified RNAs. The invention relates further to the amplification and the identification of nucleic acid molecules for the purpose of single molecule detection and/or high-throughput sequencing. In addition, a method is provided for the preparation of pooled samples that contains molecules each of which is marked by an “Identifier Sequence” for its origin. The invention facilitates studies on biological systems and analysis of genes expressed therein.
摘要翻译： 公开了用于修饰RNA分子末端的方法以及在cDNA合成中使用这些修饰的RNA分子，以克隆，检测，测序和扩增部分RNA，整个RNA或衍生的任何cDNA 来自这些修饰的RNA。 本发明进一步涉及用于单分子检测和/或高通量测序目的的核酸分子的扩增和鉴定。 此外，提供了一种用于制备包含分子的合并样品的方法，每个分子由其起源的“标识序列”标记。 本发明有助于对生物系统的研究和其中表达的基因的分析。
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公开(公告)号：US20070238158A1
公开(公告)日：2007-10-11
申请号：US11727930
申请日：2007-03-29
IPC分类号： C12N9/00
CPC分类号： C12N9/1276 , C12N9/1252 , C12N9/22 , C12N9/96
摘要： A method for enhancing activity of enzyme at an elevated temperature which comprises adding a substance exhibiting chaperone function such as a saccharide to a reaction mixture containing the enzyme. The method can improve activity of enzymes more easily and more effectively and hence afford increased enzyme activity at an elevated temperature.
摘要翻译： 一种在升高的温度下增强酶的活性的方法，其包括在含有该酶的反应混合物中加入显示伴侣功能的物质如糖。 该方法可以更容易且更有效地提高酶的活性，从而在升高的温度下提高酶的活性。
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49.发明授权公开(公告)号：US06627399B1
公开(公告)日：2003-09-30
申请号：US09622644
申请日：2000-10-23
IPC分类号： C12Q168
CPC分类号： C12Q1/6869 , C12Q2535/101 , C12Q2527/125 , C12Q2521/119
摘要： An RNA polymerase transcription accelerator comprising a compound represented by the following Formula (I) or salts thereof. A method of sequencing DNA in which nucleic acid transcripts are obtained using an RNA polymerase and a DNA fragment as a template, the resulted nucleic acid transcripts are separated, the nucleic acid sequence is determined from the separated fractions wherein the nucleic acid transcription reaction is carried out in the presence of a compound selected from a group of compounds represented by the above formula (I). The polyamine compounds above have outstanding accelerating activity on transcription activity of RNA polymerase. Therefore, use of the polyamine compounds in a DNA sequencing method using RNA polymerase can make a length of DNA sequence that can be determined in one sequencing longer.
摘要翻译： 一种包含下式（I）表示的化合物或其盐的RNA聚合酶转录促进剂。1.一种使用RNA聚合酶和DNA片段作为模板获得核酸转录物的DNA测序方法，得到的核酸转录物为 从分离的级分中确定核酸序列，其中核酸转录反应在选自上述式（I）表示的化合物的化合物的存在下进行。上述多胺化合物具有突出的加速活性 对RNA聚合酶的转录活性。 因此，在使用RNA聚合酶的DNA测序方法中使用多胺化合物可以使一段长度的DNA序列可以在一个测序中确定。
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50.发明授权Compounds having energy transfer function and method for DNA base sequencing by using the same 失效具有能量转移功能的化合物及其使用方法进行DNA碱基测序公开(公告)号：US06482938B1
公开(公告)日：2002-11-19
申请号：US09254547
申请日：1999-09-17
IPC分类号： C07H1904
CPC分类号： C07H21/00 , C07H19/10 , C07H19/20 , C12Q1/6869 , C12Q2565/1015 , C12Q2521/119
摘要： Disclosed are compounds having two kinds of reporters that can be a donor and an acceptor for energy transfer, for example, fluorescent groups, and having a 2′, 3′-dideoxyribonucleotide residue or a 3′-deoxyribonucleotide residue. These compounds can be used as terminators for the chain terminator method. The two kinds of reporters are arranged with a distance sufficient for causing energy transfer from the donor to the acceptor. Also disclosed are methods for determining DNA sequences based on the chain terminator method wherein the chain termination reaction is performed by using the above terminators. Also disclosed are compounds having two kinds of reporters that can be a donor and an acceptor for energy transfer, which can be used as a primer or an initiator in methods for determining DNA sequences utilizing the chain terminator method, and methods for determining DNA sequences utilizing the compounds.
摘要翻译： 公开了具有两种记录者的化合物，其可以是能量转移的供体和受体，例如荧光基团，并具有2'，3'-二脱氧核糖核苷酸残基或3'-脱氧核糖核苷酸残基。 这些化合物可用作链终止剂方法的终止剂。 这两种记者的排列距离足以引起从供体到受体的能量转移。 还公开了基于链终止法测定DNA序列的方法，其中通过使用上述终止子进行链终止反应。 还公开了具有两种记录者的化合物，其可以是供体和用于能量转移的受体，其可以用作引物或引发剂用于使用链终止剂法测定DNA序列的方法，以及利用 化合物。
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