RU2012149631A - СПОСОБ ПОЛУЧЕНИЯ aRNA И СПОСОБ АНАЛИЗА ЭКСПРЕССИИ ГЕНА - Google Patents
СПОСОБ ПОЛУЧЕНИЯ aRNA И СПОСОБ АНАЛИЗА ЭКСПРЕССИИ ГЕНА Download PDFInfo
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- RU2012149631A RU2012149631A RU2012149631/10A RU2012149631A RU2012149631A RU 2012149631 A RU2012149631 A RU 2012149631A RU 2012149631/10 A RU2012149631/10 A RU 2012149631/10A RU 2012149631 A RU2012149631 A RU 2012149631A RU 2012149631 A RU2012149631 A RU 2012149631A
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- arna
- utp
- uridine
- triphosphate
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1096—Processes for the isolation, preparation or purification of DNA or RNA cDNA Synthesis; Subtracted cDNA library construction, e.g. RT, RT-PCR
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- 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
- C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
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- 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
- C12Q1/6813—Hybridisation assays
- C12Q1/6827—Hybridisation assays for detection of mutation or polymorphism
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- 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
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/6865—Promoter-based amplification, e.g. nucleic acid sequence amplification [NASBA], self-sustained sequence replication [3SR] or transcription-based amplification system [TAS]
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- Life Sciences & Earth Sciences (AREA)
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- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
1. Способ получения aRNA для применения в анализе экспрессии гена с использованием образца РНК, выделенного из фиксированной ткани или клетки(клеток), включающий этап амплификации РНК, включающий этап обратной транскрипции и этап транскрипции in vitro, где доля 5-(3-аминоаллил)уридин-5'-трифосфата (AA-UTP), содержащегося в нуклеотидном реагенте, используемом на указанном этапе транскрипции in vitroсоставляет не менее 5% моль и менее 25% моль относительно общего количества уридин-5'-трифосфата (UTP) и 5-(3-аминоаллил)уридин-5'-трифосфата (AA-UTP).2. Способ получения aRNA по п.1, где указанные фиксированные ткань или клетку(клетки) заливают парафином.3. Способ получения aRNA по п.1 или 2, где средняя длина нуклеотида указанной aRNA составляет не менее 200 нуклеотидов.4. Способ анализа экспрессии гена с использованием образца РНК, выделенного из фиксированной ткани или клетки(клеток), в котором используют aRNA, полученную способом по любому из пп.1-3.5. Способ анализа по п.4, в котором используют микрочип.
Claims (5)
1. Способ получения aRNA для применения в анализе экспрессии гена с использованием образца РНК, выделенного из фиксированной ткани или клетки(клеток), включающий этап амплификации РНК, включающий этап обратной транскрипции и этап транскрипции in vitro, где доля 5-(3-аминоаллил)уридин-5'-трифосфата (AA-UTP), содержащегося в нуклеотидном реагенте, используемом на указанном этапе транскрипции in vitro, составляет не менее 5% моль и менее 25% моль относительно общего количества уридин-5'-трифосфата (UTP) и 5-(3-аминоаллил)уридин-5'-трифосфата (AA-UTP).
2. Способ получения aRNA по п.1, где указанные фиксированные ткань или клетку(клетки) заливают парафином.
3. Способ получения aRNA по п.1 или 2, где средняя длина нуклеотида указанной aRNA составляет не менее 200 нуклеотидов.
4. Способ анализа экспрессии гена с использованием образца РНК, выделенного из фиксированной ткани или клетки(клеток), в котором используют aRNA, полученную способом по любому из пп.1-3.
5. Способ анализа по п.4, в котором используют микрочип.
Applications Claiming Priority (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2010098639 | 2010-04-22 | ||
JP2010-098639 | 2010-04-22 | ||
JP2011013098 | 2011-01-25 | ||
JP2011-013098 | 2011-01-25 | ||
PCT/JP2011/059778 WO2011132726A1 (ja) | 2010-04-22 | 2011-04-21 | aRNAの調製方法および遺伝子発現の解析方法 |
Publications (1)
Publication Number | Publication Date |
---|---|
RU2012149631A true RU2012149631A (ru) | 2014-05-27 |
Family
ID=44834242
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
RU2012149631/10A RU2012149631A (ru) | 2010-04-22 | 2011-04-21 | СПОСОБ ПОЛУЧЕНИЯ aRNA И СПОСОБ АНАЛИЗА ЭКСПРЕССИИ ГЕНА |
Country Status (9)
Country | Link |
---|---|
US (1) | US20120196768A1 (ru) |
EP (1) | EP2484760B1 (ru) |
JP (1) | JP4840542B1 (ru) |
KR (1) | KR101773408B1 (ru) |
CN (1) | CN102858973A (ru) |
BR (1) | BR112012017649A2 (ru) |
CA (1) | CA2795436C (ru) |
RU (1) | RU2012149631A (ru) |
WO (1) | WO2011132726A1 (ru) |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US11091810B2 (en) * | 2015-01-27 | 2021-08-17 | BioSpyder Technologies, Inc. | Focal gene expression profiling of stained FFPE tissues with spatial correlation to morphology |
CN106198952A (zh) * | 2016-07-01 | 2016-12-07 | 清华大学 | 一种抑制核酸分子对传感界面非特异性吸附的封闭方法 |
Family Cites Families (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7229765B2 (en) * | 2000-11-28 | 2007-06-12 | Rosetta Inpharmatics Llc | Random-primed reverse transcriptase-in vitro transcription method for RNA amplification |
DK1410011T3 (da) * | 2001-06-18 | 2011-07-18 | Netherlands Cancer Inst | Diagnose og prognose for brystcancerpatienter |
US7364846B2 (en) * | 2002-10-11 | 2008-04-29 | Molecular Devices Corporation | Gene expression profiling from FFPE samples |
BRPI0414553A (pt) * | 2003-09-19 | 2006-11-07 | Arcturus Bioscience Inc | previsão do resultado de tratamento contra o cáncer de mama |
US20050196782A1 (en) * | 2003-12-23 | 2005-09-08 | Kiefer Michael C. | Universal amplification of fragmented RNA |
-
2011
- 2011-04-21 CN CN2011800201062A patent/CN102858973A/zh active Pending
- 2011-04-21 JP JP2011520487A patent/JP4840542B1/ja not_active Expired - Fee Related
- 2011-04-21 BR BR112012017649A patent/BR112012017649A2/pt not_active Application Discontinuation
- 2011-04-21 CA CA2795436A patent/CA2795436C/en not_active Expired - Fee Related
- 2011-04-21 RU RU2012149631/10A patent/RU2012149631A/ru not_active Application Discontinuation
- 2011-04-21 KR KR1020127021098A patent/KR101773408B1/ko active IP Right Grant
- 2011-04-21 US US13/499,824 patent/US20120196768A1/en not_active Abandoned
- 2011-04-21 WO PCT/JP2011/059778 patent/WO2011132726A1/ja active Application Filing
- 2011-04-21 EP EP11772058.1A patent/EP2484760B1/en not_active Not-in-force
Also Published As
Publication number | Publication date |
---|---|
EP2484760A1 (en) | 2012-08-08 |
KR20130061662A (ko) | 2013-06-11 |
WO2011132726A1 (ja) | 2011-10-27 |
KR101773408B1 (ko) | 2017-08-31 |
EP2484760A4 (en) | 2012-12-26 |
BR112012017649A2 (pt) | 2015-09-15 |
CA2795436C (en) | 2017-12-05 |
JP4840542B1 (ja) | 2011-12-21 |
EP2484760B1 (en) | 2015-10-14 |
CA2795436A1 (en) | 2011-10-27 |
JPWO2011132726A1 (ja) | 2013-07-18 |
CN102858973A (zh) | 2013-01-02 |
US20120196768A1 (en) | 2012-08-02 |
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FA93 | Acknowledgement of application withdrawn (no request for examination) |
Effective date: 20140422 |