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    • 1. 发明申请
    • ISOTHERMAL REACTIONS FOR THE AMPLIFICATION OF OLIGONUCLEOTIDES
    • 用于扩增寡核苷酸的同位素反应
    • WO2004067726A3
    • 2005-12-15
    • PCT/US2004002718
    • 2004-01-29
    • KECK GRADUATE INSTIONIAN TECHNOLOGIES INCVAN NESS JEFFREYGALAS DAVID JVAN NESS LORI KGRAYBILL JEFFREY B
    • VAN NESS JEFFREYGALAS DAVID JVAN NESS LORI KGRAYBILL JEFFREY B
    • C12N20060101C12Q1/68
    • C12Q1/6848C12Q2527/101C12Q2525/131C12Q2521/301
    • Described is a new class of isothermal reactions for amplifying DNA. These homogeneous reactions rapidly synthesize short oligonucleotides (e.g., 8-16 bases) specified by the sequence of an amplification template. Versions of the reactions can proceed in either a linear or an exponential amplification mode. Both of these reactions utilize simple, stable conditions. The rate of amplification depends entirely on the molecular parameters governing the interactions of the molecules in the reaction. The exponential version of the method is a molecular chain reaction that uses the oligonucleotide products of each linear reaction to create a producer of more of the same oligonucleotide. It is a highly sensitive chain reaction that can be triggered by specific DNA sequences and can achieve amplifications of more than 106 fold. Several similar reactions in this class are described here. The robustness, speed and sensitivity of the exponential reaction render it useful in rapidly detecting the presence of small amounts of a specific DNA sequence in a sample, and a range of other applications including many currently making use of the polymerase chain reaction (PCR).
    • 描述了一类用于扩增DNA的等温反应。 这些均相反应快速合成由扩增模板的序列指定的短寡核苷酸(例如8-16个碱基)。 反应的版本可以以线性或指数放大模式进行。 这两种反应都使用简单,稳定的条件。 扩增速率完全取决于控制反应中分子相互作用的分子参数。 该方法的指数形式是使用每个线性反应的寡核苷酸产物产生更多相同寡核苷酸的生产者的分子链反应。 它是一种高度敏感的连锁反应,可以由特定的DNA序列引发,可以达到超过106倍的扩增。 这里描述了这个类中的几个类似的反应。 指数反应的鲁棒性,速度和灵敏度使其有助于快速检测样品中少量特定DNA序列的存在,以及包括目前使用聚合酶链反应(PCR)的许多其他应用的范围。
    • 9. 发明申请
    • NUCLEIC ACID SEQUENCING USING NICKING AGENTS
    • 用酸化剂代替核酸序列
    • WO2004067764A3
    • 2005-11-10
    • PCT/US2004002719
    • 2004-01-29
    • KECK GRADUATE INSTVAN NESS JEFFREYGALAS DAVID JVAN NESS LORI K
    • VAN NESS JEFFREYGALAS DAVID JVAN NESS LORI K
    • C12Q1/68
    • C12Q1/6869C12Q1/6872C12Q2533/101C12Q2521/313C12Q2521/301C12Q2531/101C12Q2525/131
    • Nucleotide base sequence information is obtained upon combining a template nucleic acid comprising a primer strand and a target nucleic acid strand, the target nucleic acid strand comprising an interrogation sequence, with a nicking agent that recognizes a double-stranded sequence present in the template nucleic acid and then nicks the primer strand of the template nucleic acid at a location relative to the nicking agent recognition sequence, with a distributive polymerase that will extend the primer strand from the site where the nicking agent nicked the primer strand, in the presence of suitably-reactive mononucleotides, to provide a plurality of oligonucleotides that are complements of the sequence present in the interrogation sequence, where the plurality differ in having different nucleotide lengths, so the aligning the plurality of oligonucleotides provides nucleotide base sequence information about the interrogation sequence.
    • 将包含引物链和靶核酸链的模板核酸(包括询问序列的靶核酸链)与识别模板核酸中存在的双链序列的切口剂组合时获得核苷酸碱基序列信息 然后在相对于切口剂识别序列的位置处切割模板核酸的引物链,使用分布式聚合酶,其在适当的条件下存在下将延伸引物链从切口切口引物链的位点延伸, 反应性单核苷酸,以提供作为询问序列中存在的序列的互补序列的多个寡核苷酸,其中多个核苷酸长度不同,因此对齐多个寡核苷酸提供了关于询问序列的核苷酸碱基序列信息。