Automated system for polynucleotide synthesis and purification
    2.
    发明授权
    Automated system for polynucleotide synthesis and purification 失效
    用于多核苷酸合成和纯化的自动化系统

    公开(公告)号:US06175006B1

    公开(公告)日:2001-01-16

    申请号:US09281205

    申请日:1999-03-30

    IPC分类号: C07H2100

    摘要: A method and system for polynucleotide synthesis are provided which employ solid phase synthesis on a nonswellable porous polystyrene support by phosphoramidite or hydrogen phosphonate chemistries. The polystyrene support gives rise to fewer tritylated failure sequences caused by chain growth from extraneous support sites, and allows lower amounts of monomer reactants to be used to achieve equal or better coupling efficiencies as those achieveable with CPG. The method and system also employ nucleoside intermediates whose exocyclic amines are protected by base-labile groups which permit simultaneous cleavage and deprotection of the completed polynucleotide chain in the presence of the solid phase support. This latter feature allows practical automation of both the synthesis and purification of polynucleotides.

    摘要翻译: 提供了一种用于多核苷酸合成的方法和系统,其通过亚磷酰胺或氢化膦酸盐化学物质在不可分散的多孔聚苯乙烯载体上进行固相合成。 聚苯乙烯支持物产生由来自外部支持位点的链增长引起的三苯甲基化失败序列的更少,并且允许使用较少量的单体反应物来实现与CPG可达到的相同或更好的偶联效率。 该方法和系统还使用核苷中间体,其外环胺由碱不稳定基团保护,其允许在固相载体存在下同时裂解和脱保护完整的多核苷酸链。 后一个特征允许多核苷酸的合成和纯化的实际自动化。

    Reversible di-nucleotide terminator sequencing
    5.
    发明授权
    Reversible di-nucleotide terminator sequencing 有权
    可逆二核苷酸终止子测序

    公开(公告)号:US08715938B2

    公开(公告)日:2014-05-06

    申请号:US13198901

    申请日:2011-08-05

    申请人: Gerald Zon

    发明人: Gerald Zon

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

    摘要: The present teachings provide methods, compositions, and kits for synthesizing and sequencing nucleic acids. In some embodiments, reversible di-nucleotide compounds are employed along with cleaving reactions that remove a label and a blocking moiety. Improved sequencing efficiency is achieved by the rapid polymerase-mediated incorporation of reversible di-nucleotide compounds. In some embodiments, the di-nucleotides do not contain conventional nucleotide triphosphates, but rather employ amino acid phosphoramidate nucleotides (AAPNs).

    摘要翻译: 本教导提供了用于合成和测序核酸的方法,组合物和试剂盒。 在一些实施方案中,使用可逆的二核苷酸化合物以及去除标记物和阻断部分的切割反应。 通过快速聚合酶介导的可逆二核苷酸化合物的掺入可以改善测序效率。 在一些实施方案中,二核苷酸不含常规核苷酸三磷酸,而是使用氨基酸氨基磷酸核苷酸(AAPN)。

    CHEMICALLY MODIFIED LIGASE COFACTORS, DONORS AND ACCEPTORS
    6.
    发明申请
    CHEMICALLY MODIFIED LIGASE COFACTORS, DONORS AND ACCEPTORS 有权
    化学改性助剂,助剂和接收剂

    公开(公告)号:US20110008788A1

    公开(公告)日:2011-01-13

    申请号:US12831212

    申请日:2010-07-06

    IPC分类号: C12Q1/68 C12N9/00 C12P19/34

    摘要: Provided herein are methods for ligation of polynucleotides containing modified ligation components, particularly modified ligase cofactors, modified acceptors and modified donors. The methods readily applied to ligation-based assays for detection of a nucleic acid sequence where the use of the modified cofactor improves discrimination between matched and mismatched templates. Furthermore, the use of the modified ligation components reduces or eliminates the ligation in the absence of nucleic acid template.

    摘要翻译: 本文提供了用于连接含有修饰的连接组分,特别是修饰的连接酶辅因子,修饰的受体和修饰的供体的多核苷酸的方法。 该方法容易地应用于用于检测核酸序列的基于连接的测定,其中使用经修饰的辅因子改善了匹配和不匹配模板之间的区别。 此外,修饰的连接组分的使用减少或消除了在没有核酸模板的情况下的连接。

    Method of forming N-protected amino acid thiohydantoins
    9.
    发明授权
    Method of forming N-protected amino acid thiohydantoins 失效
    形成N-保护的氨基酸硫代乙内酰脲的方法

    公开(公告)号:US5304497A

    公开(公告)日:1994-04-19

    申请号:US914280

    申请日:1992-07-15

    摘要: A method of forming a thiohydantoin from an N-protected amino acid. The method employs a uronium or phosphonium compound to activate the terminal carboxyl group of the amino acid and a thiocyanate reagent to cyclize the activated amino acid to the thio-hydantoin. The thiohydantoin can be cleaved from its N-protecting group, for use in C-terminal peptide sequencing. Particularly preferred uronium compounds include salts of 2-chlorouronium. Preferred thiocyanate reagents include trimethylsilyl isothiocyanate and crown ether adducts of metallothiocyanates, such as the 18-crown-6 adduct of KSCN.

    摘要翻译: 从N-保护的氨基酸形成硫代乙内酰脲的方法。 该方法采用铀或鏻化合物活化氨基酸的末端羧基和硫氰酸酯试剂,使活化的氨基酸环化成硫代乙内酰脲。 硫代乙内酰脲可以从其N-保护基裂解,用于C末端肽测序。 特别优选的尿素化合物包括2-氯脲的盐。 优选的硫氰酸酯试剂包括异硫氰酸三甲基甲硅烷基酯和金属硫氰酸酯的冠醚加合物,例如KSCN的18-冠-6加合物。

    Method and Materials for Quaternary Amine Catalyzed Bisulfite Conversion of Cytosine to Uracil
    10.
    发明申请
    Method and Materials for Quaternary Amine Catalyzed Bisulfite Conversion of Cytosine to Uracil 审中-公开
    第四纪胺催化亚硫酸氢盐将胞嘧啶转化成尿嘧啶的方法和材料

    公开(公告)号:US20100087641A1

    公开(公告)日:2010-04-08

    申请号:US12468820

    申请日:2009-05-19

    IPC分类号: C07D239/02 C12N9/12 B01J31/02

    摘要: The invention provides methods and materials for the conversion of cytosine to uracil. A nucleic acid, such a gDNA, is reacted with bisulfate, such as magnesium bisulfite, in the presence of a quaternary amine catalyst. Examples of suitable quaternary amine catalysts include but are not limited to quaternary ammonium compounds, quaternary alkyl ammonium salts, quaternary alkyl ammonium halides, quaternary methyl ammonium bromide, quaternary ammonium chloride, tetraethyl ammonium hydroxide, tetraethylammonium chloride, tetrabutyl ammonium chloride, tetrabutyl ammonium bromide. The invention also contemplates kits of premeasured ingredients for carrying out the methods of the invention either on an individual sample or on a plurality of samples.

    摘要翻译: 本发明提供了将胞嘧啶转化成尿嘧啶的方法和材料。 在四胺催化剂存在下,使核酸,例如gDNA与硫酸氢盐如亚硫酸氢镁反应。 合适的季胺催化剂的实例包括但不限于季铵化合物,季烷基铵盐,季烷基卤化铵,季铵甲基溴化铵,季铵氯化物,四乙基氢氧化铵,四乙基氯化铵,四丁基氯化铵,四丁基溴化铵。 本发明还考虑了用于在单个样品或多个样品上进行本发明的方法的预测成分的试剂盒。