公开(公告)号：US06218118B1

公开(公告)日：2001-04-17

申请号：US09112437

申请日：1998-07-09

申请人： Jeffrey R. Sampson ,  Zohar H. Yakhini ,  Peter G. Webb ,  Nicholas M. Sampas ,  Anna M. Tsalenko ,  Joel Myerson

发明人： Jeffrey R. Sampson ,  Zohar H. Yakhini ,  Peter G. Webb ,  Nicholas M. Sampas ,  Anna M. Tsalenko ,  Joel Myerson

IPC分类号： C12Q168

CPC分类号： C12Q1/6872 ,  B01J2219/00454 ,  B01J2219/00497 ,  B01J2219/00605 ,  B01J2219/0061 ,  B01J2219/00612 ,  B01J2219/00626 ,  B01J2219/00637 ,  B01J2219/00641 ,  B01J2219/00722 ,  C12Q1/6874 ,  C40B40/00 ,  C12Q2537/143 ,  C12Q2533/101 ,  C12Q2533/107

摘要： Methods and reagents are disclosed which satisfy the need for more sensitive, more accurate and higher through-put analyses of target nucleic acid sequences. The methods and reagents may be generically applied to generally any target nucleic acid sequence and do not require a priori information about the presence, location or identity of mutations in the target nucleic acid sequence. The reagents of the invention are mixtures of natural and mass-modified oligonucleotide precursors having a high level of coverage and mass number complexity. A method is also disclosed for analyzing a target nucleic acid sequence employing the mixtures of natural and mass-modified oligonucleotide precursors and chemical or enzymatic assays to alter the mass of the oligonucleotide precursors prior to mass spectral analysis, generally via MALDI-TOF. The enzymatic assay may be a polymerase extension assay or a ligase assay. The kits for carrying out the methods of the invention are also disclosed.

摘要翻译： 公开了满足对靶核酸序列的更灵敏，更准确和更高的通过分析的需要的方法和试剂。 方法和试剂通常可以一般地应用于任何靶核酸序列，并且不需要关于靶核酸序列中突变的存在，位置或特征的先验信息。 本发明的试剂是具有高覆盖率和质量数复杂性的天然和质量修饰的寡核苷酸前体的混合物。 还公开了一种使用天然和质量修饰寡核苷酸前体的混合物分析目标核酸序列的方法，以及通常通过MALDI-TOF在质谱分析之前改变寡核苷酸前体的质量以进行化学或酶学测定。 酶测定可以是聚合酶延伸测定或连接酶测定。 还公开了用于实施本发明的方法的试剂盒。

公开(公告)号：US07399844B2

公开(公告)日：2008-07-15

申请号：US09836012

申请日：2001-04-17

申请人： Jeffrey R. Sampson ,  Joel Myerson ,  Anna M. Tsalenko ,  Nicholas M. Sampas ,  Peter G. Webb ,  Zohar H. Yakhini

发明人： Jeffrey R. Sampson ,  Joel Myerson ,  Anna M. Tsalenko ,  Nicholas M. Sampas ,  Peter G. Webb ,  Zohar H. Yakhini

IPC分类号： C07H19/00 ,  C12Q1/68

CPC分类号： C12Q1/6872 ,  B01J2219/00452 ,  B01J2219/00454 ,  B01J2219/00497 ,  B01J2219/00527 ,  B01J2219/00581 ,  B01J2219/00585 ,  B01J2219/00596 ,  B01J2219/00599 ,  B01J2219/00605 ,  B01J2219/0061 ,  B01J2219/00612 ,  B01J2219/00626 ,  B01J2219/00637 ,  B01J2219/00641 ,  B01J2219/00659 ,  B01J2219/00707 ,  B01J2219/00722 ,  C12Q1/6874 ,  C12Q2600/156 ,  C40B40/00 ,  C12Q2537/143 ,  C12Q2533/101 ,  C12Q2533/107

摘要： Methods and reagents are disclosed which provide for more sensitive, more accurate and higher through-put analyses of target nucleic acid sequences. The methods and reagents of the present invention may be generically applied to generally any target nucleic acid sequence and do not require a priori information about the presence, location or identity of mutations in the target nucleic acid sequence. The reagents of the invention are mixtures of oligonucleotide precursors having a high level of coverage and mass number complexity, and also having tags analyzable by mass spectrometry which are covalently linked to the precursors through cleavable bonds. A method is also disclosed for analyzing a target nucleic acid sequence employing the mixtures of oligonucleotide precursors having tags analyzable by mass spectrometry covalently linked to the oligonucleotide precursors through cleavable bonds, and chemical or enzymatic assays to alter the mass of the oligonucleotide precursors prior to mass spectral analysis. The enzymatic assay may be a polymerase extension assay or a ligation-based assay. The kits for carrying out the methods of the invention are also disclosed.

摘要翻译： 公开了提供对靶核酸序列的更灵敏，更准确和更高的通过分析的方法和试剂。 本发明的方法和试剂通常可以一般地应用于任何靶核酸序列，并且不需要关于靶核酸序列中突变的存在，位置或同一性的先验信息。 本发明的试剂是具有高水平的覆盖率和质量数复杂性的寡核苷酸前体的混合物，并且还具有可通过质谱法分析的标签，其通过可切割键与前体共价连接。 还公开了一种用于分析靶核酸序列的方法，该寡核苷酸前体具有通过可通过可切割键与寡核苷酸前体共价连接的质谱分析的标签的寡核苷酸前体的混合物，以及在质量之前改变寡核苷酸前体的质量的化学或酶学测定 光谱分析。 酶测定可以是聚合酶延伸测定或基于连接的测定。 还公开了用于实施本发明的方法的试剂盒。