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    DNA sequencing by mass spectrometry
    25.
    发明授权
    DNA sequencing by mass spectrometry 失效
    通过质谱法进行DNA测序

    公开(公告)号：US5547835A

    公开(公告)日：1996-08-20

    申请号：US178216

    申请日：1994-01-06

    Applicant: Hubert Koster

    Inventor： Hubert Koster

    IPC: C12N15/09 ,  C07H19/20 ,  C07H21/04 ,  C12P19/34 ,  C12Q1/68 ,  G01N27/62 ,  G01N33/483 ,  G01N33/50 ,  G01N35/10 ,  C07H21/02

    CPC classification number: C12Q1/6827 ,  C12Q1/6816 ,  C12Q1/6872 ,  G01N35/1067 ,  Y10S435/81 ,  Y10T436/143333 ,  Y10T436/24 ,  Y10T436/25

    Abstract: The invention describes a new method to sequence DNA. The improvements over the existing DNA sequencing technologies are high speed, high throughput, no electrophoresis and gel reading artifacts due to the complete absence of an electrophoretic step, and no costly reagents involving various substitutions with stable isotopes. The invention utilizes the Sanger sequencing strategy and assembles the sequence information by analysis of the nested fragments obtained by base-specific chain termination via their different molecular masses using mass spectrometry, as for example, MALDI or ES mass spectrometry. A further increase in throughput can be obtained by introducing mass-modifications in the oligonucleotide primer, chain-terminating nucleoside triphosphates and/or in the chain-elongating nucleoside triphosphates, as well as using integrated tag sequences which allow multiplexing by hybridization of tag specific probes with mass differentiated molecular weights.

    Abstract translation: 本发明描述了一种新的DNA序列化方法。 由于完全不存在电泳步骤，现有DNA测序技术的改进是高速度，高通量，无电泳和凝胶阅读伪像，并且没有昂贵的试剂涉及具有稳定同位素的各种取代。 本发明利用Sanger测序策略，并通过使用质谱法（例如MALDI或ES质谱法）通过分析通过其不同分子量通过碱基特异性链终止获得的嵌套片段来组装序列信息。 通过在寡核苷酸引物，链终止的三磷酸核苷和/或在延伸链的三磷酸的链中引入质量修饰，以及使用允许通过标签特异性探针的杂交进行多重复合的整合的标签序列，可以获得通量的进一步提高 具有质量差异分子量。

    Process for the preparation of oligonucleotides
    26.
    发明授权
    Process for the preparation of oligonucleotides 失效
    寡核苷酸的制备方法

    公开(公告)号：US4725677A

    公开(公告)日：1988-02-16

    申请号：US752178

    申请日：1985-06-18

    Applicant: Hubert Koster ,  Nanda D. Sinha

    Inventor： Hubert Koster ,  Nanda D. Sinha

    IPC: C07H19/067 ,  C07F9/26 ,  C07H19/073 ,  C07H19/10 ,  C07H19/20 ,  C07H21/00 ,  C07H21/04 ,  C07H15/12 ,  C07H17/00

    CPC classification number: C07H21/00 ,  C07F9/26 ,  Y02P20/55

    Abstract: The invention relates to a process for the preparation of oligonucleotides by the following steps: reaction of a nucleoside with a phosphine derivative, reaction of the nucleotide derivative thus obtained with a nucleoside bonded to a polymeric carrier, oxidation of the carrier-bound nucleoside-nucleotide thus obtained with formation of phosphotriester groups, blocking of free primary 5'-OH groups, elimination of a protective group from the terminal 5'-OH group, where appropriate single or multiple repetition of the abovementioned steps to introduce further nucleoside phosphate or oligonucleoside phosphate units, and cleavage of the nucleoside-carrier bond and, where appropriate, elimination of all protective groups present in the oligonucleoside phosphates. The phosphine derivative used is a compound of the general formula III ##STR1## in which X and L can react with OH groups of the sugar units in the oligonucleotides, and R.sup.3 is a protective group which can be liberated by .beta.-elimination.

    Abstract translation: PCT No.PCT / EP84 / 00244 Sec。 371日期：1985年6月18日 102（e）日期1985年6月18日PCT提交1984年8月10日PCT公布。 出版物WO85 / 0081600 日本1985年2月28日。本发明涉及通过以下步骤制备寡核苷酸的方法：核苷与膦衍生物的反应，由此得到的核苷酸衍生物与与聚合物载体结合的核苷的反应，氧化 由此形成磷酸三酯基团的载体结合的核苷 - 核苷酸，阻断游离的原代5'-OH基团，从末端5'-OH基团消除保护基团，其中适当的单次或多次重复上述步骤 引入另外的磷酸核苷或磷酸核苷单元，并切割核苷 - 载体键，并在适当情况下消除寡核苷酸磷酸酯中存在的所有保护基。 所使用的膦衍生物是通式III（III）的化合物，其中X和L可以与寡核苷酸中的糖单元的OH基反应，R3是可以通过β-消光释放的保护基 。

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