RU2016107198A - Аналоги нуклеотидов - Google Patents

Аналоги нуклеотидов Download PDF

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RU2016107198A
RU2016107198A RU2016107198A RU2016107198A RU2016107198A RU 2016107198 A RU2016107198 A RU 2016107198A RU 2016107198 A RU2016107198 A RU 2016107198A RU 2016107198 A RU2016107198 A RU 2016107198A RU 2016107198 A RU2016107198 A RU 2016107198A
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nucleotide
nucleic acid
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propargyl
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RU2016107198A
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Дэ Хюнь КИМ
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Эбботт Молекьюлар Инк.
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    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
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    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
    • C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
    • C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
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    • C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
    • C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
    • C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
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    • C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
    • C07H21/04—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids with deoxyribosyl as saccharide radical
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    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1034—Isolating an individual clone by screening libraries
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    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1034—Isolating an individual clone by screening libraries
    • C12N15/1093—General methods of preparing gene libraries, not provided for in other subgroups
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    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/26—Preparation of nitrogen-containing carbohydrates
    • C12P19/28—N-glycosides
    • C12P19/30—Nucleotides
    • C12P19/34—Polynucleotides, e.g. nucleic acids, oligoribonucleotides
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6869—Methods for sequencing

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  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Saccharide Compounds (AREA)

Claims (19)

1. Композиция, содержащая аналог нуклеотида, имеющий структуру в соответствии с:
Figure 00000001
где В представляет собой основание, и Р содержит фосфатную группу.
2. Композиция по п. 1, отличающаяся тем, что Р содержит тетрафосфат, трифосфат, дифосфат, монофосфат, модифицированный тетрафосфат, модифицированный трифосфат, модифицированный дифосфат или модифицированный монофосфат.
3. Композиция по п. 1, отличающаяся тем, что В выбран из группы, состоящей из цитозина, гуанина, аденина, тимина и урацила.
4. Композиция по п. 1, отличающаяся тем, что В содержит пурин, пиримидин, модифицированный пурин или модифицированный пиримидин.
5. Композиция по п. 1, отличающаяся тем, что указанный аналог нуклеотида содержит тиоалкинил, тиопропаргил, 3'-S-пропаргил, тиофуранозу, тиорибозу, тиодезоксирибозу, арабинозу или модифицированный сахар.
6. Композиция по п. 1, отличающаяся тем, что Ρ содержит 5'-гидроксил, альфа-тиофосфат, бета-тиофосфат, гамма-тиофосфат, альфа-метилфосфонат, бета-метилфосфонат или гамма-метилфосфонат.
7. Способ секвенирования нуклеиновой кислоты, при этом указанный способ включает:
a) гибридизацию праймера с матрицей нуклеиновой кислоты с образованием комплекса гибридизованного праймера/матрицы нуклеиновой кислоты;
b) обеспечение множества аналогов нуклеотидов, при этом каждый аналог нуклеотида содержит алкиновую группу;
c) проведение реакции указанного комплекса гибридизованного праймера/матрицы нуклеиновой кислоты и указанного аналога нуклеотида с полимеразой с присоединением указанного аналога нуклеотида к указанному праймеру путем полимеразной реакции с получением удлиненного продукта, содержащего встроенный аналог нуклеотида; и
d) проведение реакции указанного удлиненного продукта с азидсодержащим соединением с получением структуры, содержащей триазольное кольцо.
8. Способ по п. 7, отличающийся тем, что указанные аналоги нуклеотидов представляют собой 3'-О-пропаргил-дНТФ-аналоги нуклеотидов, и Η выбран из группы, состоящей из А, С, G, Τ и U.
9. Способ по п. 7, отличающийся тем, что указанную структуру, содержащую триазольное кольцо, применяют в последующих ферментативных реакциях.
10. Способ по п. 9, отличающийся тем, что указанная последующая ферментативная реакция представляет собой полимеразную цепную реакцию.
11. Способ по п. 7, включающий обеспечение стандартных нуклеотидов.
12. Библиотека NGS, содержащая множество нуклеиновых кислот, при этом каждая нуклеиновая кислота содержит 3'-О-пропаргильный нуклеотид.
13. Реакционная смесь, содержащая один или более дНТФ и один или более 3'-О-пропаргильных аналогов нуклеотидов.
RU2016107198A 2013-08-19 2014-08-19 Набор для получения реакционной смеси для синтеза 3′-o-пропаргил-модифицированной нуклеиновой кислоты RU2688435C2 (ru)

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US201361867202P 2013-08-19 2013-08-19
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US20150051113A1 (en) 2015-02-19
CN105722850A (zh) 2016-06-29
EP3036248A4 (en) 2017-03-01
ES2805004T3 (es) 2021-02-10
US20210207192A1 (en) 2021-07-08
CA2921619C (en) 2021-05-25
EP3036248A2 (en) 2016-06-29
US9932623B2 (en) 2018-04-03
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US20200157606A1 (en) 2020-05-21
RU2688435C2 (ru) 2019-05-21
CA3112661A1 (en) 2015-02-26
US20150050697A1 (en) 2015-02-19
RU2016107198A3 (ru) 2018-08-01
CA2921619A1 (en) 2015-02-26
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