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91. 发明授权

US5962228A DNA extension and analysis with rolling primers 失效
标题翻译： DNA扩增和分析与滚动引物
公开(公告)号：US5962228A
公开(公告)日：1999-10-05
申请号：US916120
申请日：1997-08-22
申请人： Sydney Brenner
发明人： Sydney Brenner
IPC分类号： C12Q1/68 , C07H21/02 , C07H21/04 , C12N15/00
CPC分类号： C12Q1/6874 , C12Q1/6869 , B01J2219/00648 , B01J2219/00659
摘要： A novel "primer walking" method for DNA sequencing is provided that comprises repeated cycles nucleotide identification by selective extension and primer advancement along a template by template mutation. An important feature of the invention is providing a set of primers, referred to herein as "rolling primers" that contain complexity-reducing nucleotides for reducing the number of primers required for annealing to every possible primer binding site on a sequencing template. Another important feature of the invention is the systematic replacement of at least one of the four nucleotides in the target polynucleotide with its cognate complexity-reducing nucleotide or complement thereof. Sequencing is initiated by annealing rolling primers differing only in their terminal nucleotides to a primer binding site of a sequencing template so that only the rolling primer whose terminal nucleotide forms a perfect complement with the template leads to the formation of an extension product. After amplifying the double stranded extension product to form an amplicon, the terminal nucleotide, and hence its complement in the template, is identified by the identity of the amplicon. The primer binding site of the template of the successfully amplified polynucleotide is then mutated by, for example, oligonucleotide-directed mutagenesis so that a subsequent rolling primer may be selected from the set that forms a perfectly matched duplex with the mutated template at a site which is shifted towards the direction of extension by one nucleotide relative to the binding site of the previous rolling primer. The steps of selective extension, amplification and identification are then repeated. In this manner, the primers "roll" along the polynucleotide during the sequencing process, moving a base at a time along the template with each cycle.
摘要翻译：提供了一种用于DNA测序的新型“引物步行”方法，其包括通过模板突变沿着模板的选择性延伸和引物前进重复循环核苷酸鉴定。本发明的一个重要特征是提供一组引物，在本文中称为“滚动引物”，其含有复杂度降低的核苷酸，用于将退火所需的引物数量减少到测序模板上的每个可能的引物结合位点。本发明的另一个重要特征是用其同源复杂度降低的核苷酸或其互补体系系统地置换靶多核苷酸中的四个核苷酸中的至少一个。通过将仅在其末端核苷酸不同的滚动引物退火到测序模板的引物结合位点来启动测序，使得只有末端核苷酸与模板完全互补的滚动引物导致扩增产物的形成。在扩增双链延伸产物以形成扩增子后，终端核苷酸，因此其模板中的补体由扩增子的鉴定鉴定。然后通过例如寡核苷酸定向诱变将成功扩增的多核苷酸的模板的引物结合位点突变，使得随后的翻转引物可以选自与突变模板形成完全匹配的双链体的组，相对于先前的滚动底漆的结合位点向延伸方向移动一个核苷酸。然后重复选择性扩增，扩增和鉴定步骤。以这种方式，引物在测序过程中沿着多核苷酸“滚动”，每个周期沿模板一次移动碱基。

92. 发明授权

US5863722A Method of sorting polynucleotides 失效
标题翻译：多核苷酸排序方法
公开(公告)号：US5863722A
公开(公告)日：1999-01-26
申请号：US485105
申请日：1995-06-07
申请人： Sydney Brenner
发明人： Sydney Brenner
IPC分类号： G01N33/53 , A61K49/00 , B01J19/00 , C07H21/00 , C07H21/04 , C12N15/09 , C12N15/10 , C12N15/11 , C12Q1/68 , C40B40/06 , C40B70/00 , G01N33/566 , G01N37/00
CPC分类号： B01J19/0046 , C07H21/00 , C12N15/10 , C12N15/1034 , C12N15/1065 , C12Q1/6809 , C12Q1/6813 , C12Q1/6827 , C12Q1/6834 , C12Q1/6839 , C12Q1/6855 , C12Q1/6869 , C12Q1/6874 , C40B80/00 , B01J2219/005 , B01J2219/00572 , B01J2219/00605 , B01J2219/0061 , B01J2219/00612 , B01J2219/00626 , B01J2219/0063 , B01J2219/00637 , B01J2219/00648 , B01J2219/00659 , B01J2219/00722 , C40B40/06 , C40B70/00
摘要： The invention provides a method and materials for sorting polynucleotides with oligonucleotide tags. Oligonucleotide tags of the invention are capable of hybridizing to complementary oligomeric compounds consisting of subunits having enhanced binding strength and specificity as compared to natural oligonucleotides. Such complementary oligomeric compounds are referred to herein as "tag complements." Subunits of tag complements may consist of monomers of non-natural nucleotide analogs, referred to herein as "antisense monomers" or they may comprise oligomers having lengths in the range of 3 to 6 nucleotides or analogs thereof, including antisense monomers, the oligomers being selected from a minimally cross-hybridizing set. In such a set, a duplex made up of an oligomer of the set and the complement of any other oligomer of the set contains at least two mismatches. Preferred antisense monomers include peptide nucleic acid monomers and nucleoside phosphoramidates having a 3'--NHP(O)(O--)O--5' linkage with its adjacent nucleoside.An important aspect of the invention is the use of the oligonucleotide tags for sorting polynucleotides by specifically hybridizing tags attached to the polynucleotides to their complements on solid phase supports. This embodiment provides a readily automated system for manipulating and sorting polynucleotides, particularly useful in large-scale parallel operations, such as large-scale DNA sequencing, mRNA fingerprinting, or the like, wherein many target polynucleotides or many segments of a single target polynucleotide are sequenced simultaneously.
摘要翻译：本发明提供了用寡核苷酸标签分选多核苷酸的方法和材料。与天然寡核苷酸相比，本发明的寡核苷酸标签能够与由具有增强的结合强度和特异性的亚基组成的互补寡聚化合物杂交。这种互补寡聚化合物在本文中称为“标签互补物”。标签互补子单元可以由非天然核苷酸类似物的单体组成，本文称为“反义单体”，或者它们可以包含长度在3至6个核苷酸范围内的寡核苷酸或其类似物，包括反义单体，所选择的寡聚体从最小交叉杂交集。在这样的组中，由该组的低聚物和该组的任何其它低聚物的互补体组成的双链体包含至少两个错配。优选的反义单体包括与其相邻核苷具有3'-NHP（O）（O）O-5'键的肽核酸单体和核苷氨基磷酸酯。本发明的一个重要方面是使用寡核苷酸标签通过将与多核苷酸连接的标签与其固相载体上的互补序列特异性杂交来分选多核苷酸。该实施方案提供了用于操纵和分选多核苷酸的容易自动化的系统，特别可用于大规模并行操作，例如大规模DNA测序，mRNA指纹图谱等，其中单个靶多核苷酸的许多靶多核苷酸或许多片段是同时排序。

93. 发明授权

US5654413A Compositions for sorting polynucleotides 失效
标题翻译：用于分选多核苷酸的组合物
公开(公告)号：US5654413A
公开(公告)日：1997-08-05
申请号：US484712
申请日：1995-06-07
申请人： Sydney Brenner
发明人： Sydney Brenner
IPC分类号： C12N15/09 , A61K49/00 , B01J19/00 , C07H20060101 , C07H21/00 , C12N20060101 , C12N15/10 , C12N15/63 , C12P19/34 , C12Q20060101 , C12Q1/00 , C12Q1/68 , C40B40/06 , C40B70/00 , G01N33/53 , G01N33/566 , G01N37/00
CPC分类号： B01J19/0046 , C07H21/00 , C12N15/10 , C12N15/1034 , C12N15/1065 , C12Q1/6809 , C12Q1/6816 , C12Q1/6827 , C12Q1/6834 , C12Q1/6837 , C12Q1/6855 , C12Q1/6874 , C40B80/00 , B01J2219/005 , B01J2219/00572 , B01J2219/00605 , B01J2219/0061 , B01J2219/00612 , B01J2219/00626 , B01J2219/0063 , B01J2219/00637 , B01J2219/00641 , B01J2219/00648 , B01J2219/00659 , B01J2219/00722 , C40B40/06 , C40B70/00
摘要： The invention provides a method of tracking, identifying, and/or sorting classes or subpopulations of molecules by the use of oligonucleotide tags. Oligonucleotide tags of the invention each consist of a plurality of subunits 3 to 6 nucleotides in length selected from a minimally cross-hybridizing set. A subunit of a minimally cross-hybridizing set forms a duplex or triplex having two or more mismatches with the complement of any other subunit of the same set. The number of oligonucleotide tags available in a particular embodiment depends on the number of subunits per tag and on the length of the subunit. An important aspect of the invention is the use of the oligonucleotide tags for sorting polynucleotides by specifically hybridizing tags attached to the polynucleotides to their complements on solid phase supports. This embodiment provides a readily automated system for manipulating and sorting polynucleotides, particularly useful in large-scale parallel operations, such as large-scale DNA sequencing, mRNA fingerprinting, and the like, wherein many target polynucleotides or many segments of a single target polynucleotide are sequenced simultaneously.
摘要翻译：本发明提供了通过使用寡核苷酸标签来跟踪，鉴定和/或分类分子的分类或亚群的方法。本发明的寡核苷酸标签每个由选自最小交叉杂交组的长度为3至6个核苷酸的多个亚单位组成。最小交叉杂交组的亚基形成具有两个或更多个错配的双链体或三链体，同一组的任何其他亚基的互补体。在特定实施方案中可用的寡核苷酸标签的数量取决于每个标签的亚单位的数目和子单元的长度。本发明的一个重要方面是使用寡核苷酸标签通过将与多核苷酸连接的标签与其固相载体上的互补序列特异性杂交来分选多核苷酸。该实施方案提供了用于操纵和分选多核苷酸的容易自动化系统，特别可用于大规模并行操作，例如大规模DNA测序，mRNA指纹图谱等，其中单个靶多核苷酸的许多靶多核苷酸或多个片段是同时排序。

94. 发明授权

US5635400A Minimally cross-hybridizing sets of oligonucleotide tags 失效
标题翻译：微小交叉杂交寡核苷酸标签组
公开(公告)号：US5635400A
公开(公告)日：1997-06-03
申请号：US478238
申请日：1995-06-07
申请人： Sydney Brenner
发明人： Sydney Brenner
IPC分类号： C12N15/09 , A61K49/00 , B01J19/00 , C07H20060101 , C07H21/00 , C12N20060101 , C12N15/10 , C12N15/63 , C12P19/34 , C12Q20060101 , C12Q1/00 , C12Q1/68 , C40B40/06 , C40B70/00 , G01N33/53 , G01N33/566 , G01N37/00
CPC分类号： B01J19/0046 , C07H21/00 , C12N15/10 , C12N15/1034 , C12N15/1065 , C12Q1/6809 , C12Q1/6816 , C12Q1/6827 , C12Q1/6834 , C12Q1/6837 , C12Q1/6855 , C12Q1/6874 , C40B80/00 , B01J2219/005 , B01J2219/00572 , B01J2219/00605 , B01J2219/0061 , B01J2219/00612 , B01J2219/00626 , B01J2219/0063 , B01J2219/00637 , B01J2219/00641 , B01J2219/00648 , B01J2219/00659 , B01J2219/00722 , C40B40/06 , C40B70/00
摘要： The invention provides a method of tracking, identifying, and/or sorting classes or subpopulations of molecules by the use of oligonucleotide tags. Oligonucleotide tags of the invention each consist of a plurality of subunits 3 to 6 nucleotides in length selected from a minimally cross-hybridizing set. A subunit of a minimally cross-hybridizing set forms a duplex or triplex having two or more mismatches with the complement of any other subunit of the same set. The number of oligonucleotide tags available in a particular embodiment depends on the number of subunits per tag and on the length of the subunit. An important aspect of the invention is the use of the oligonucleotide tags for sorting polynucleotides by specifically hybridizing tags attached to the polynucleotides to their complements on solid phase supports. This embodiment provides a readily automated system for manipulating and sorting polynucleotides, particularly useful in large-scale parallel operations, such as large-scale DNA sequencing, mRNA fingerprinting, and the like, wherein many target polynucleotides or many segments of a single target polynucleotide are sequenced simultaneously.
摘要翻译：本发明提供了通过使用寡核苷酸标签来跟踪，鉴定和/或分类分子的分类或亚群的方法。本发明的寡核苷酸标签每个由选自最小交叉杂交组的长度为3至6个核苷酸的多个亚单位组成。最小交叉杂交组的亚基形成具有两个或更多个错配的双链体或三链体，同一组的任何其他亚基的互补体。在特定实施方案中可用的寡核苷酸标签的数量取决于每个标签的亚单位的数目和子单元的长度。本发明的一个重要方面是使用寡核苷酸标签通过将与多核苷酸连接的标签与其固相载体上的互补序列特异性杂交来分选多核苷酸。该实施方案提供了用于操纵和分选多核苷酸的容易自动化系统，特别可用于大规模并行操作，例如大规模DNA测序，mRNA指纹图谱等，其中单个靶多核苷酸的许多靶多核苷酸或多个片段是同时排序。

95. 发明授权

US5573905A Encoded combinatorial chemical libraries 失效
公开(公告)号：US5573905A
公开(公告)日：1996-11-12
申请号：US860445
申请日：1992-03-30
申请人： Richard Lerner , Kim Janda , Sydney Brenner
发明人： Richard Lerner , Kim Janda , Sydney Brenner
IPC分类号： G01N33/53 , C07H21/00 , C07H21/04 , C07K14/00 , C12N15/09 , C12N15/10 , C12Q1/68 , C40B70/00
CPC分类号： C12N15/1075 , C07H21/00 , C12N15/1062 , B01J2219/00572 , C40B70/00
摘要： The present invention describes an encoded combinatorial chemical library comprised of a plurality of bifunctional molecules having both a chemical polymer and an identifier oligonucleotide sequence that defines the structure of the chemical polymer. Also described are the bifunctional molecules of the library, and methods of using the library to identify chemical structures within the library that bind to biologically active molecules in preselected binding interactions.

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