RU2000104843A - Анализ последовательности нуклеиновой кислоты - Google Patents
Анализ последовательности нуклеиновой кислотыInfo
- Publication number
- RU2000104843A RU2000104843A RU2000104843/13A RU2000104843A RU2000104843A RU 2000104843 A RU2000104843 A RU 2000104843A RU 2000104843/13 A RU2000104843/13 A RU 2000104843/13A RU 2000104843 A RU2000104843 A RU 2000104843A RU 2000104843 A RU2000104843 A RU 2000104843A
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Claims (29)
1. Способ секвенирования полинуклеотида, включающий этапы:
(a) реагирование полинуклеотида-мишени с полимеразой, иммобилизованной на твердой подложке, и различными нуклеотидами в условиях, обеспечивающих полимеразную реакцию;
(b) детекция проявления последствий инкорпорации конкретного нуклеотида, комплементарного полинуклеотиду-мишени.
(a) реагирование полинуклеотида-мишени с полимеразой, иммобилизованной на твердой подложке, и различными нуклеотидами в условиях, обеспечивающих полимеразную реакцию;
(b) детекция проявления последствий инкорпорации конкретного нуклеотида, комплементарного полинуклеотиду-мишени.
2. Способ по п. 1, отличающийся тем, что проявление по этапу (b) выявляется путем измерения излучения.
3. Способ по п. 1 или 2, отличающийся тем, что этапы (а) и (b) осуществляются с каждым из различных нуклеотидов по очереди до того момента, как происходит выявление инкорпорации, с последующим повтором этих этапов.
4. Способ по п. 1 или 2, отличающийся тем, что этап (а) осуществляется в присутствии всех нуклеотидов.
5. Способ по любому из предыдущих пунктов, отличающийся тем, что нуклеотиды. включают 3'-блокирующую группу, которую удаляют после прохождения реакции полимеризации.
6. Способ по п. 5, отличающийся тем, что блокирующая группа может быть избирательно удалена воздействием пульсирующего монохроматического света.
7. Способ по п. 5 или 6, отличающийся тем, что нуклеотиды включают дополнительную блокирующую группу по концевому фосфатному остатку в трифосфатной цепочке, а дополнительная блокирующая группа удаляется до удаления 3'-блокирующей группы.
8. Способ по п. 7, отличающийся тем, что дополнительная блокирующая группа может быть избирательно удалена действием пульсирующего монохроматического света в условиях, отличающихся от тех, которые необходимы для удаления 3'-блокирующей группы.
9. Способ по п. 8, отличающийся тем, что дополнительная блокирующая группа удаляется действием пульсирующего монохроматического света в течение времени отличного от требуемого для удаления 3'-блокирующей группы.
10. Способ по любому из предыдущих пунктов, отличающийся тем, что этап (а) дополнительно включает внесение конкурентного ингибитора полимеразы.
11. Способ по любому из предыдущих пунктов, отличающийся тем, что полинуклеотид-мишень на этапе (а) связан с полимеразой за счет β--димерного комплекса.
12. Способ по любому из предыдущих пунктов, отличающийся тем, что полимеразой является ДНК-полимераза III E. coli или полимераза Т7.
13. Способ по любому из пп. 1-11, отличающийся тем, что полимеразой является Таg-полимераза.
14. Способ по любому из пп. 1-11, отличающийся тем, что полимеразой является обратная транскриптаза.
15. Способ по любому из пп. 1-14, отличающийся тем, что этап (b) включает выявление изменений резонансного сигнала во времени.
16. Способ по любому из пп. 1-15, отличающийся тем, что излучение является электромагнитным.
17. Способ по п. 16, отличающийся тем, что электромагнитное излучение относится к инфракрасной области спектра.
18. Способ по любому из пп. 1-17, отличающийся тем, что этап (b) включает применение резонанса плазмонов на поверхности.
19. Способ по п. 16, отличающийся тем, что электромагнитное излучение относится к спектру радиочастот.
20. Способ по п. 19, отличающийся тем, что инкорпорация нуклеотида определяется с применением ядерно-магнитного резонанса.
21. Способ по любому из пп. 1-20, отличающийся тем, что полинуклеотидом является ДНК.
22. Сенсорный чип, несущий иммобилизованную на него полимеразу.
23. Нуклеотид, включающий блокирующую группу по его 3'-положению и по концевому фосфатному остатку трифосфатной цепочки, при том, что эти две блокирующие группы могут быть удалены действием монохроматического света с разной длиной волны.
24. Нуклеотид по п. 23, отличающийся тем, что блокирующая группа является производной от соединения формулы R1-[О-СО-] Х, где R1 представлен фотолабильной группой, а Х представлен отщепляемой группой.
25. Нуклеотид по п. 23 или 24, отличающийся тем, что блокирующей группой по 3'-положению является о-нитробензилоксикарбонильная группа.
26. Нуклеотид по любому из пп. 23-25, отличающийся тем, что блокирующей группой по концевому фосфату является о-нитробензильная группа.
27. Нуклеотид по любому из пп. 23-26, отличающийся тем, что блокирующей группой по 3'-положению является (4,5-диметокси-2-нитробензил) оксикарбонильная группа.
28. Нуклеотид по любому из пп. 23-27, отличающийся тем, что блокирующей группой по концевому фосфату является 1-(2-нитрофенил)этиловая группа.
29. Устройство для секвенирования полинуклеотида, состоящее из оптического сенсорного чипа, источника света, устройства считывания изображения и светочувствительного элемента, отличающийся тем, что сенсорный чип представляет собой чип по п. 22.
Applications Claiming Priority (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| GBGB9715942.0A GB9715942D0 (ru) | 1997-07-28 | 1997-07-28 | |
| GB9715942.0 | 1997-07-28 | ||
| GB9727103.5 | 1997-12-22 | ||
| GBGB9727103.5A GB9727103D0 (ru) | 1997-12-22 | 1997-12-22 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| RU2000104843A true RU2000104843A (ru) | 2002-03-10 |
| RU2198221C2 RU2198221C2 (ru) | 2003-02-10 |
Family
ID=26311958
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| RU2000104843/13A RU2198221C2 (ru) | 1997-07-28 | 1998-07-24 | Способ секвенирования полинуклеотида и устройство для его осуществления |
Country Status (18)
| Country | Link |
|---|---|
| US (4) | US7008766B1 (ru) |
| EP (4) | EP2267165B1 (ru) |
| JP (1) | JP2001511358A (ru) |
| KR (1) | KR100664331B1 (ru) |
| CN (1) | CN1152140C (ru) |
| AT (1) | ATE225858T1 (ru) |
| AU (1) | AU735898B2 (ru) |
| BR (1) | BR9812270A (ru) |
| CA (1) | CA2294962C (ru) |
| DE (1) | DE69808661T3 (ru) |
| DK (1) | DK1017848T3 (ru) |
| ES (1) | ES2183394T5 (ru) |
| IL (1) | IL133411A (ru) |
| IS (1) | IS5360A (ru) |
| NZ (1) | NZ502557A (ru) |
| PT (1) | PT1017848E (ru) |
| RU (1) | RU2198221C2 (ru) |
| WO (1) | WO1999005315A2 (ru) |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| RU2334958C2 (ru) * | 2003-04-04 | 2008-09-27 | Интернэшнл Фронтьер Сайенс Организейшн | Устройство и способ для улучшенного наноспектроскопического сканирования |
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