公开(公告)号：US06235478B1

公开(公告)日：2001-05-22

申请号：US09287682

申请日：1999-04-06

Applicant: Hubert Köster

Inventor： Hubert Köster

IPC: C12Q168

CPC classification number: C12Q1/6872 ,  C12Q1/6816 ,  C12Q1/6827 ,  C12Q1/6837 ,  C12Q1/6858 ,  C12Q1/686 ,  C12Q1/6862 ,  C12Q1/6883 ,  C12Q1/706 ,  G01N35/1067 ,  H01J49/00 ,  Y10T436/143333 ,  Y10T436/24 ,  C12Q2565/627 ,  C12Q2537/113 ,  C12Q2535/113 ,  C12Q2521/325 ,  C12Q2535/131 ,  C12Q2525/107 ,  C12Q2523/107 ,  C12Q2523/319 ,  C12Q2565/625 ,  C12Q2535/125

Abstract: Fast and highly accurate mass spectrometry-based processes for detecting particular nucleic acid molecules and sequences in the molecules are provided. Depending upon the sequence to be detected, the processes, for example, can be used to diagnose a genetic disease or a chromosomal abnormality, a predisposition to a disease or condition, or infection by a pathogen, or for determining identity or heredity.

公开(公告)号：US06225450B1

公开(公告)日：2001-05-01

申请号：US08481033

申请日：1995-06-07

Applicant: Hubert Köster

Inventor： Hubert Köster

IPC: C12Q163

CPC classification number: C12Q1/6827 ,  C12Q1/6816 ,  C12Q1/6872 ,  G01N35/1067 ,  Y10S435/81 ,  Y10T436/143333 ,  Y10T436/24 ,  Y10T436/25 ,  C12Q2565/627 ,  C12Q2563/167 ,  C12Q2537/113 ,  C12Q2537/143

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 flirter 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 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。 可以通过在寡核苷酸引物，链终止的三磷酸核苷和/或延伸链的三磷酸的链中引入质量修饰，以及使用允许通过杂交标记特异性探针多重化的综合标签序列来获得通过量的调节剂增加， 质量分辨分子量。