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RU2019138698A - Способы и композиции для днк-профилирования - Google Patents

Способы и композиции для днк-профилирования Download PDF

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RU2019138698A
RU2019138698A RU2019138698A RU2019138698A RU2019138698A RU 2019138698 A RU2019138698 A RU 2019138698A RU 2019138698 A RU2019138698 A RU 2019138698A RU 2019138698 A RU2019138698 A RU 2019138698A RU 2019138698 A RU2019138698 A RU 2019138698A
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nucleic acid
acid sample
primers
snp
amplification
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RU2019138698A
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RU2752700C2 (ru
RU2019138698A3 (ru
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Кэтрин М. СТЕФЕНС
Сидне ХОЛТ
Кэри ДЭВИС
Анне ЯГЕР
Паулина ВАЛИХЕВИЧ
Йонми ХАН
Дэвид СИЛВА
Минь-Цзюи Ричард ШЭНЬ
Сасан АМИНИ
Фрэнк СТИМЕРС
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Иллумина, Инк.
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Claims (23)

1. Способ получения ДНК–профиля, включающий
обеспечение образца нуклеиновой кислоты,
амплификацию образца нуклеиновой кислоты с множеством праймеров, которые специфически гибридизуются по меньшей мере с одной последовательностью-мишенью, содержащей однонуклеотидный полиморфизм (SNP), и по меньшей мере с одной последовательностью-мишенью, содержащей тандемный повтор, в мультиплексной реакции с получением продуктов амплификации, где по меньшей мере один из множества праймеров имеет низкую температуру плавления, имеет длину по меньшей мере 24 нуклеотида или содержит гомополимерную нуклеотидную последовательность, или их комбинацию, и
определение в продуктах амплификации генотипов по меньшей мере одного SNP и по меньшей мере одного тандемного повтора, таким образом, получая ДНК-профиль образца нуклеиновой кислоты.
2. Способ по п.1, в котором образец нуклеиновой кислоты получают у человека.
3. Способ по п.1, в котором по меньшей мере один SNP включает SNP, который, как известно, указывает на родственную или фенотипическую характеристику источника образца нуклеиновой кислоты.
4. Способ по п. 1, в котором по меньшей мере один из множества праймеров содержит по меньшей мере 24 нуклеотида.
5. Способ по п.4, в котором по меньшей мере один из множества праймеров имеет температуру плавления менее 60°С.
6. Способ по п.1, в котором амплификация образца нуклеиновой кислоты включает проведение полимеразной цепной реакции (ПЦР) на образце нуклеиновой кислоты.
7. Способ по п.1, в котором множество праймеров специфически гибридизуются по меньшей мере с 30 SNP.
8. Способ по п.1, в котором множество праймеров специфически гибридизуются по меньшей мере с 24 последовательностями тандемных повторов.
9. Способ по п.1, в котором образец нуклеиновой кислоты содержит геномную ДНК.
10. Способ по п.9, в котором обеспечение образца нуклеиновой кислоты включает обеспечение криминалистического образца, содержащего нуклеиновую кислоту.
11. Способ по п.1, в котором определение генотипов включает определение по меньшей мере 90% генотипов по меньшей мере одного SNP и по меньшей мере одного тандемного повтора.
12. Способ по п.1, в котором, по меньшей мере, один из множества праймеров содержит одну или несколько последовательностей–меток, содержащих метку уникального молекулярного идентификатора.
13. Способ построения библиотеки нуклеиновых кислот, включающий: предоставление образца нуклеиновой кислоты и амплификацию образца нуклеиновой кислоты со множеством праймеров, которые специфически гибридизуются с по меньшей мере одним однонуклеотидным полиморфизмом (SNP) и по меньшей мере с одной последовательностью-мишенью, содержащей последовательность тандемного повтора в мультиплексной реакции для получения продуктов амплификации, где по меньшей мере один из множества праймеров имеет низкую температуру плавления, имеет длину по меньшей мере 24 нуклеотида или содержит гомополимерную нуклеотидную последовательность, или их комбинацию.
14. Способ по п.13, в котором по меньшей мере один SNP указывает на родственную или фенотипическую характеристику источника образца нуклеиновой кислоты.
15. Способ по п.13, в котором по меньшей мере один из множества праймеров содержит одну или несколько последовательностей-меток.
16. Способ по п.15, в котором одна или несколько последовательностей-меток содержат метку-праймер, метку для захвата, метку для секвенирования или метку уникального молекулярного идентификатора или их сочетание.
17. Способ по п.16, дополнительно включающий амплификацию продуктов амплификации со вторым множеством праймеров.
18. Способ по п.17, в котором по меньшей мере один из второго множества праймеров содержит часть, соответствующую метке-праймеру из множества праймеров и одной или нескольким последовательностям-меткам.
19. Способ по п.17, включающий добавление связывающего одноцепочечные ДНК белка (SSB) к продуктам амплификации.
20. Способ по п.13, в котором амплификация образца нуклеиновой кислоты включает амплификацию посредством полимеразной цепной реакции (ПЦР).
RU2019138698A 2014-02-18 2015-02-13 Способы и композиции для днк-профилирования RU2752700C2 (ru)

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US201462043060P 2014-08-28 2014-08-28
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