公开(公告)号：US5885775A

公开(公告)日：1999-03-23

申请号：US726090

申请日：1996-10-04

申请人： Lawrence A. Haff ,  Igor Pavlovich Smirnov

发明人： Lawrence A. Haff ,  Igor Pavlovich Smirnov

IPC分类号： C12Q1/68 ,  C12P19/34 ,  G01N24/00

CPC分类号： C12Q1/6886 ,  C12Q1/6858 ,  C12Q2600/16 ,  Y10T436/24

摘要： The invention relates to methods for determining sequence information in polynucleotides by combining the recent disparate technologies of mass spectrometry and polynucleotide hybridization, amplification, extension and/or ligation techniques. Broadly, in a first step, the method for determining sequence information in a sample polynucleotide includes hybridizing with a sample nucleotide one or a mixture of oligonucleotide probes having a nucleotide sequence complementary to a portion of the sample polynucleotide, thereby forming a complex. Then, in a second step, the complex is contacted with at least a member selected from the group consisting of nucleosides, dideoxynucleosides, polymerases, nucleases, transcriptases, ligases and restriction enzymes to alter at least a subset of said oligonucleotide probes. In a third step, the method provides for determining the molecular weight of at least the subset of altered probes by mass spectrometry and infering the sequence information of the sample polynucleotide therefrom.

摘要翻译： 本发明涉及通过组合最近不同的质谱技术和多核苷酸杂交，扩增，延伸和/或连接技术来确定多核苷酸序列信息的方法。 广泛地，在第一步中，用于确定样品多核苷酸中的序列信息的方法包括与样品核苷酸1或具有与样品多核苷酸的一部分互补的核苷酸序列的寡核苷酸探针的混合物杂交，从而形成复合物。 然后，在第二步中，将复合物与至少一种选自核苷，双脱氧核苷，聚合酶，核酸酶，转录酶，连接酶和限制酶的成员接触以改变至少一个所述寡核苷酸探针的子集。 在第三步中，所述方法提供通过质谱法确定至少所述改变的探针子集的分子量，并从其中推断样品多核苷酸的序列信息。