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91. 发明申请

US20060211003A1 One component and two component DNA Pol lll replicases and uses thereof 有权
标题翻译：一个组分和两个组分的DNA Pol lll复制酶及其用途
公开(公告)号：US20060211003A1
公开(公告)日：2006-09-21
申请号：US11101977
申请日：2005-04-07
申请人： Lars-Erik Peters , Anna Ettinger
发明人： Lars-Erik Peters , Anna Ettinger
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6867 , C12N9/1252 , C12Q2521/101
摘要： The invention provides one-component and two-component DNA polymerases, as well as kits comprising the same, and methods of using the same for nucleic amplification and nucleic acid sequencing.
摘要翻译：本发明提供单组分和双组分DNA聚合酶，以及包含其的试剂盒，以及将其用于核酸扩增和核酸测序的方法。

92. 发明申请

US20060172309A1 5-methylcytosine detection, compositions and methods therefor 有权
标题翻译： 5-甲基胞嘧啶检测，其组合物和方法
公开(公告)号：US20060172309A1
公开(公告)日：2006-08-03
申请号：US10881363
申请日：2004-06-30
申请人： Gerald Zon
发明人： Gerald Zon
IPC分类号： C12Q1/68 , C07J17/00 , C07H19/04
CPC分类号： C12Q1/6827 , C07H19/16 , C12Q1/6869 , C12Q2565/501 , C12Q2563/107 , C12Q2521/101
摘要： Compositions and methods for detecting 5-methylcytosine in a nucleic acid are disclosed. A 5-methylcytosine discriminator, which is a deoxyribonucleosidetriphosphate comprising a cytosine-pairing moiety such as a guanosine and a moiety which hinders hydrogen bonding between the cytosine-pairing moiety and a 5-methylcytosine is described. The discriminator is able to base pair with a cytosine but not a 5-methylcytosine. A 5-methylcytosine comprised by a target nucleotide can be detected in a reaction using a DNA polymerase and a primer hybridized immediately adjacent to the target nucleotide. In the reaction, pyrophosphate released upon incorporation of a dNTP complementary to a target nucleotide is detected. Lack of incorporation of the discriminator, but incorporation of a dGTP, can indicate that the target nucleotide is a 5-methylcytosine.
摘要翻译：公开了用于检测核酸中5-甲基胞嘧啶的组合物和方法。描述了一种5-甲基胞嘧啶鉴别剂，其是包含胞嘧啶配对部分如鸟苷和妨碍胞嘧啶配对部分与5-甲基胞嘧啶之间的氢键的部分的脱氧核糖核苷三磷酸。鉴别器能够与胞嘧啶碱基配对，但不能与5-甲基胞嘧啶碱基配对。可以使用DNA聚合酶和与靶核苷酸紧邻杂交的引物在反应中检测由靶核苷酸组成的5-甲基胞嘧啶。在反应中，检测到掺入与目标核苷酸互补的dNTP时释放的焦磷酸盐。缺乏鉴别器的并入，但掺入dGTP可以表明目标核苷酸是5-甲基胞嘧啶。

93. 发明申请

US20060088877A1 Systematic evolution of ligands by exponential enrichment: Chemi-SELEX 审中-公开
标题翻译：配体通过指数富集的系统演化：Chemi-SELEX
公开(公告)号：US20060088877A1
公开(公告)日：2006-04-27
申请号：US11296832
申请日：2005-12-06
申请人： Larry Gold , Bruce Eaton , Drew Smith , Matthew Wecker , Kirk Jensen
发明人： Larry Gold , Bruce Eaton , Drew Smith , Matthew Wecker , Kirk Jensen
IPC分类号： C12Q1/68 , C12P19/34 , C07H21/04
CPC分类号： C12N15/115 , A61K47/547 , A61K47/549 , B82Y5/00 , C07H19/06 , C07H19/10 , C07H21/00 , C07K14/001 , C12N9/1276 , C12N15/1048 , C12N2310/13 , C12N2310/322 , C12N2310/53 , C12P19/34 , C12Q1/37 , C12Q1/6804 , C12Q1/6811 , C12Q1/6876 , C12Q1/70 , C40B40/00 , F02B2075/027 , G01N33/531 , G01N33/532 , G01N33/535 , G01N33/56988 , G01N33/68 , G01N33/76 , G01N2333/163 , G01N2333/575 , G01N2333/62 , G01N2333/8125 , G01N2333/96433 , G01N2333/96455 , G01N2333/966 , G01N2333/9726 , G01N2333/974 , G01N2333/976 , C12Q2541/101 , C12Q2525/101 , C12Q2525/179 , C12Q2525/155 , C12Q2521/101
摘要： This application provides methods for identifying nucleic acid ligands capable of covalently interacting with targets of interest. The nucleic acids can be associated with various functional units. The method also allows for the identification of nucleic acids that have facilitating activities as measured by their ability to facilitate formation of a covalent bond between the nucleic acid, including its associated functional unit, and its target.
摘要翻译：该应用提供了用于鉴定能够与感兴趣的靶标共价相互作用的核酸配体的方法。核酸可以与各种功能单元相关联。该方法还允许鉴定具有促进活性的核酸，其通过其促进在包括其相关功能单位的核酸与其靶标之间形成共价键的能力来测量。

94. 发明申请

US20060084099A1 Product identification method 审中-公开
标题翻译：产品识别方法
公开(公告)号：US20060084099A1
公开(公告)日：2006-04-20
申请号：US11237884
申请日：2005-09-29
申请人： Hiroshi Yokoyama , Masahiko Yamanaka , Kentarou Watanabe , Tsunehiko Higuchi , Hidetoshi Hayashi
发明人： Hiroshi Yokoyama , Masahiko Yamanaka , Kentarou Watanabe , Tsunehiko Higuchi , Hidetoshi Hayashi
IPC分类号： C12Q1/68 , G06F19/00 , C12P19/34
CPC分类号： C12Q1/6816 , G16B30/00 , C12Q2563/185 , C12Q2521/101
摘要： A product identification method of one embodiment of the invention includes the steps of providing a product with an information nucleic acid having a segment of known base sequence randomly selected for the product, extracting the information nucleic acid from the product, amplifying the information nucleic acid by a polymerase chain reaction; and determining a base sequence of the information nucleic acid to identify the product by the base sequence of the information nucleic acid.
摘要翻译：本发明的一个实施方案的产品鉴定方法包括以下步骤：向产品提供具有随机选择用于产品的已知碱基序列的信息核酸，从产物中提取信息核酸，通过以下步骤扩增信息核酸：聚合酶链反应; 以及根据所述信息核酸的碱基序列确定所述信息核酸的碱基序列以鉴定所述产物。

95. 发明申请

US20060084074A1 Use of whole blood in PCR reactions 有权
公开(公告)号：US20060084074A1
公开(公告)日：2006-04-20
申请号：US11005559
申请日：2004-12-06
申请人： Milko Kermekchiev , Wayne Barnes
发明人： Milko Kermekchiev , Wayne Barnes
IPC分类号： C12Q1/68 , C12P19/34
CPC分类号： C12N9/1252 , C12Q1/686 , C12Q2549/101 , C12Q2521/101
摘要： A method of obtaining DNA amplification of a nucleic acid target from a volume of whole blood comprising performing DNA amplification in a PCR assay mixture with a blood-resistant polymerase.

96. 发明申请

US20060057592A1 Methods for identifying primase trinucleotide initiation sites and identification of inhibitors of primase activity 审中-公开
标题翻译：用于鉴定引物启动子三核苷酸引发位点的方法和鉴定启动子酶活性抑制剂
公开(公告)号：US20060057592A1
公开(公告)日：2006-03-16
申请号：US10941717
申请日：2004-09-15
申请人： Mark Griep , Steven Hinrichs , Scott Koepsell , Khalid Sayood , James Takacs
发明人： Mark Griep , Steven Hinrichs , Scott Koepsell , Khalid Sayood , James Takacs
IPC分类号： C12Q1/68
CPC分类号： C12Q1/25 , C12Q1/68 , G01N2500/00 , Y02A50/57 , C12Q2521/101
摘要： Methods and kits for the identification of a primase trinucleotide initiation site and for the identification of compounds which modulate bacterial primase activity are provided.
摘要翻译：提供了用于鉴定起始酶三核苷酸引发位点和鉴定调节细菌引发酶活性的化合物的方法和试剂盒。

97. 发明申请

US20050255563A1 Method for plasmid preparation by conversion of open circular plasmid to supercoiled plasmid 审中-公开
标题翻译：通过将开环质粒转化为超螺旋质粒的质粒制备方法
公开(公告)号：US20050255563A1
公开(公告)日：2005-11-17
申请号：US10947360
申请日：2004-09-23
申请人： Edward Hyman
发明人： Edward Hyman
IPC分类号： C12P19/34 , C12Q1/68
CPC分类号： C12Q1/6806 , C12Q2521/113 , C12Q2521/501 , C12Q2521/101
摘要： In one embodiment of the invention, a method is provided for preparing plasmid from host cells which contain the plasmid, comprising: (a) providing a plasmid solution comprised of unligatable open circular plasmid; (b) reacting the unligatable open circular plasmid with one or more enzymes and appropriate nucleotide cofactors, such that unligatable open circular plasmid is converted to 3′-hydroxyl, 5′-phosphate nicked plasmid; (c) reacting the 3′-hydroxyl, 5′-phosphate nicked plasmid with a DNA ligase and DNA ligase nucleotide cofactor, such that 3′-hydroxyl, 5′-phosphate nicked plasmid is converted to relaxed covalently closed circular plasmid; and (d) reacting the relaxed covalently closed circular plasmid with a DNA gyrase and DNA gyrase nucleotide cofactor, such that relaxed covalently closed circular plasmid is converted to negatively supercoiled plasmid. In other embodiments, DNA gyrase is replaced with reverse DNA gyrase or reaction (d) is not performed.
摘要翻译：在本发明的一个实施方案中，提供了一种用于从含有质粒的宿主细胞制备质粒的方法，包括：（a）提供由未结合的开环质粒构成的质粒溶液; （b）使未结合的开放环状质粒与一种或多种酶和适当的核苷酸辅因子反应，使得将无重复的开环质粒转化为3'-羟基，5'-磷酸切口质粒; （c）使3'-羟基，5'-磷酸酯切口质粒与DNA连接酶和DNA连接酶核苷酸辅因子反应，使3'-羟基，5'-磷酸切口质粒转化为松弛共价闭合的环状质粒; 和（d）使松弛的共价闭合的环状质粒与DNA促旋酶和DNA促旋酶核苷酸辅因子反应，使得松弛的共价闭合的环状质粒转化为负的超螺旋质粒。在其他实施方案中，DNA逆转酶用逆DNA回旋酶代替，或不进行反应（d）。

98. 发明申请

US20050255499A1 Process for discrimination of nucleic acid sequence variants 失效
标题翻译：鉴别核酸序列变体的方法
公开(公告)号：US20050255499A1
公开(公告)日：2005-11-17
申请号：US11043099
申请日：2005-01-27
申请人： David Gelfand , Pamela Holland , Randall Saiki , Robert Watson
发明人： David Gelfand , Pamela Holland , Randall Saiki , Robert Watson
IPC分类号： C07H21/00 , C12N15/09 , C12Q1/68 , C12Q1/70
CPC分类号： C12Q1/686 , C07H21/00 , C12Q1/6818 , C12Q1/6823 , C12Q1/6827 , C12Q1/703 , Y10S435/805 , Y10S435/81 , Y10S436/815 , C12Q2521/101 , C12Q2565/1015 , C12Q2531/113 , C12Q2521/319 , C12Q2535/131 , C12Q2521/325 , C12Q2537/157 , C12Q2561/101 , C12Q2525/186 , C12Q2525/131
摘要： A process of detecting a target nucleic acid using labeled oligonucleotides uses the 5′ to 3′ nuclease activity of a nucleic acid polymerase to cleave annealed labeled oligonucleotide from hybridized duplexes and release labeled oligonucleotide fragments for detection. This process is easily incorporated into a PCR amplification assay.
摘要翻译：使用标记的寡核苷酸检测靶核酸的过程使用核酸聚合酶的5'至3'核酸酶活性从杂交双链体切割退火标记的寡核苷酸，并释放标记的寡核苷酸片段进行检测。该方法容易地并入PCR扩增测定中。

99. 发明申请

US20050089910A1 DNA polymerase having ability to reduce innate selective discrimination against fluorescent dye-labeled dideoxynucleotides 审中-公开
标题翻译： DNA聚合酶具有减少荧光染料标记的双脱氧核苷酸的先天性选择性鉴别的能力
公开(公告)号：US20050089910A1
公开(公告)日：2005-04-28
申请号：US10959015
申请日：2004-10-05
申请人： GuoFan Hong , Wei-hua Huang
发明人： GuoFan Hong , Wei-hua Huang
IPC分类号： C12N1/21 , C12N9/12 , C12Q1/68 , C07H21/04 , C12N9/22 , C12N15/74
CPC分类号： C12N9/1252 , C12Q1/6869 , C12Q2563/107 , C12Q2535/101 , C12Q2521/101
摘要： The invention relates to genetical modification of DNA polymerase to reduce its innate selective sequence-related discrimination against incorporation of fluorescent dye-labeled ddCTP and ddATP in the enzymatic reaction for preparation of samples for automated florescent dye-labeled terminator DNA sequencing. The modified DNA polymerases are more resistant to heat inactivation and are more effective in dideoxynucleotide incorporation than current DNA polymerases.
摘要翻译：本发明涉及DNA聚合酶的遗传修饰，以减少其在用于制备用于自动荧光染料标记的终止子DNA测序的样品的酶反应中掺入荧光染料标记的ddCTP和ddATP的先天选择性序列相关鉴别。修饰的DNA聚合酶对热灭活更具抗性，并且在双脱氧核苷酸掺入中比目前的DNA聚合酶更有效。

100. 发明申请

US20050069887A1 Dna synthesis promoters dna polymerase-associated factors and utilization thereof 有权
标题翻译： Dna合成启动子dna聚合酶相关因子及其利用
公开(公告)号：US20050069887A1
公开(公告)日：2005-03-31
申请号：US10495581
申请日：2002-11-14
申请人： Masao Kitabayashi , Toshihiro Kuroita , Yoshikazu Ishida , Hideyuki Komatsubara , Yoshiaki Nishiya , Masanori Oka , Yoshihisa Kawamura , Tadayuki Imanaka
发明人： Masao Kitabayashi , Toshihiro Kuroita , Yoshikazu Ishida , Hideyuki Komatsubara , Yoshiaki Nishiya , Masanori Oka , Yoshihisa Kawamura , Tadayuki Imanaka
IPC分类号： C07K14/195 , C12N9/12 , C12P19/34 , C12Q1/68
CPC分类号： C07K14/195 , C12N9/1252 , C12Q1/6846 , C12Q2527/125 , C12Q2521/101
摘要： A composition for enhancing synthesis of DNA comprising an anion-supplying substance that is effective in promoting DNA synthesis in enzymatic DNA synthesis reactions, in particular a salt of a dicarboxylic acid. Further enhanced DNA synthesis promoting effects can be achieved by using, together with the anion-supplying substance, at least one compound selected from the group consisting of dimethylsulfoxide and compounds represented by the following formula (1): R2—CH2—NR1xHy (1) wherein R1 is an alkyl group having 1 to 3 carbon atoms, R2 is a substituent selected from the group consisting of the following (a) and (b): (a) oxygen and (b) radicals represented by the formula wherein R4 is methyl, hydrogen or forms a pyrrolidine ring when combined with R1, R5 is —CO2H or —SO3H, and n is an integer from 0 to 2, x is an integer from 1 to 3 and y is an integer from 0 to 2, provided that x plus y equals 3. The present invention further provides a thermostable DNA polymerase-related factor derived from Thermococcus species, which has an activity to promote the DNA synthesis activity of DNA polymerase or bind to DNA polymerase.
摘要翻译：一种用于增强DNA合成的组合物，其包含在促进DNA合成反应中有效促进DNA合成的阴离子供给物质，特别是二羧酸的盐。通过与阴离子供给物质一起使用至少一种选自二甲基亚砜和下式（1）表示的化合物的化合物，可以实现进一步增强的DNA合成促进效果：R 2 -CH 2 -NR <1> xHy（1）其中R 1是具有1至3个碳原子的烷基，R 2是选自以下（a）和（b）的取代基：（a）氧和（b）由下式表示的基团其中R 4是甲基，氢或当与R 1一起形成吡咯烷环时，R 5是-CO 2 H或-SO 3 H，n是0至 2，x是1至3的整数，y是0至2的整数，条件是x加y等于3.本发明还提供了源自Thermococcus物种的热稳定DNA聚合酶相关因子，其具有活性促进DNA聚合酶的DNA合成活性或与DNA聚合酶结合。

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