EA200100490A1 - METHOD OF INDEXATION AND DETERMINATION OF RELATIVE CONCENTRATION OF EXPRESSING INFORMATION RNA - Google Patents

METHOD OF INDEXATION AND DETERMINATION OF RELATIVE CONCENTRATION OF EXPRESSING INFORMATION RNA

Info

Publication number
EA200100490A1
EA200100490A1 EA200100490A EA200100490A EA200100490A1 EA 200100490 A1 EA200100490 A1 EA 200100490A1 EA 200100490 A EA200100490 A EA 200100490A EA 200100490 A EA200100490 A EA 200100490A EA 200100490 A1 EA200100490 A1 EA 200100490A1
Authority
EA
Eurasian Patent Office
Prior art keywords
mrna
cdna
sequence
crna
primers
Prior art date
Application number
EA200100490A
Other languages
Russian (ru)
Inventor
Карл В. Хасел
Брайан С. Хилбуш
Original Assignee
Диджитал Гене Текнолоджиз, Инк.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Диджитал Гене Текнолоджиз, Инк. filed Critical Диджитал Гене Текнолоджиз, Инк.
Publication of EA200100490A1 publication Critical patent/EA200100490A1/en

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Classifications

    • 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
    • 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
    • C12Q1/6809—Methods for determination or identification of nucleic acids involving differential detection

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Analytical Chemistry (AREA)
  • Microbiology (AREA)
  • Immunology (AREA)
  • Molecular Biology (AREA)
  • Biotechnology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

Усовершенствованный метод одновременной последовательность-специфической идентификации мРНК в популяции мРНК позволяет визуализировать почти каждую мРНК, экспрессирующуюся тканью, в виде отдельной полосы на геле, интенсивность которой приблизительно соответствует концентрации этой мРНК. Вообще метод включает образование кДНК с использованием заякоривающих праймеров для фиксации 3'-го конца, получение клонированных вставок из этой кДНК в векторе, содержащем бактериофаг-специфический промотор для последующего синтеза РНК, получение линеаризованных фрагментов этих клонированных вставок, приготовление кРНК, транскрибирование кДНК с кРНК и осуществление двух последовательность-специфичных ПЦР-амплификаций этой кДНК. В предпочтительных реализациях этот метод включает сравнение длины и, по крайней мере, части нуклеотидной последовательности продуктов ПЦР с ожидаемыми величинами, определенными из базы данных нуклеотидных последовательностей. Этот метод может идентифицировать изменения в экспрессии мРНК, связанные с приемом (введением) лекарственных препаратов или с физиологическими или патологическими условиями. Кроме того, представляются векторы и праймеры, полезные для осуществления на практике этого усовершенствованного метода.Международная заявка была опубликована вместе с отчетом о международном поиске.The improved method for the simultaneous sequence-specific identification of mRNA in an mRNA population allows visualizing almost every mRNA expressed by a tissue as a separate band on a gel, the intensity of which approximately corresponds to the concentration of this mRNA. In general, the method involves the formation of cDNA using anchoring primers for fixing the 3'th end, obtaining cloned inserts from this cDNA in a vector containing a bacteriophage-specific promoter for subsequent synthesis of RNA, obtaining linearized fragments of these cloned inserts, preparing cRNA, transcribing cDNA with cRNA and the implementation of two sequence-specific PCR amplifications of this cDNA. In preferred implementations, this method includes comparing the length and at least part of the nucleotide sequence of the PCR products with the expected values determined from the database of nucleotide sequences. This method can identify changes in mRNA expression associated with drug intake (administration) or with physiological or pathological conditions. In addition, vectors and primers are presented that are useful for practicing this improved method. The international application was published along with an international search report.

EA200100490A 1998-11-04 1999-10-14 METHOD OF INDEXATION AND DETERMINATION OF RELATIVE CONCENTRATION OF EXPRESSING INFORMATION RNA EA200100490A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US18686998A 1998-11-04 1998-11-04
PCT/US1999/023655 WO2000026406A1 (en) 1998-11-04 1999-10-14 METHOD FOR INDEXING AND DETERMINING THE RELATIVE CONCENTRATION OF EXPRESSED MESSENGER RNAs

Publications (1)

Publication Number Publication Date
EA200100490A1 true EA200100490A1 (en) 2001-10-22

Family

ID=22686605

Family Applications (1)

Application Number Title Priority Date Filing Date
EA200100490A EA200100490A1 (en) 1998-11-04 1999-10-14 METHOD OF INDEXATION AND DETERMINATION OF RELATIVE CONCENTRATION OF EXPRESSING INFORMATION RNA

Country Status (11)

Country Link
EP (1) EP1127159A1 (en)
JP (1) JP2002528135A (en)
KR (1) KR20010092721A (en)
CN (1) CN1331755A (en)
AU (1) AU1108900A (en)
CA (1) CA2350168A1 (en)
EA (1) EA200100490A1 (en)
IL (1) IL142965A0 (en)
MX (1) MXPA01004550A (en)
NO (1) NO20012203L (en)
WO (1) WO2000026406A1 (en)

Families Citing this family (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6096503A (en) * 1993-11-12 2000-08-01 The Scripps Research Institute Method for simultaneous identification of differentially expresses mRNAs and measurement of relative concentrations
US6110680A (en) * 1993-11-12 2000-08-29 The Scripps Research Institute Method for simultaneous identification of differentially expressed mRNAs and measurement of relative concentrations
US9777312B2 (en) 2001-06-30 2017-10-03 Enzo Life Sciences, Inc. Dual polarity analysis of nucleic acids
US20040161741A1 (en) 2001-06-30 2004-08-19 Elazar Rabani Novel compositions and processes for analyte detection, quantification and amplification
US9261460B2 (en) 2002-03-12 2016-02-16 Enzo Life Sciences, Inc. Real-time nucleic acid detection processes and compositions
US20030215839A1 (en) * 2002-01-29 2003-11-20 Peter Lonnerberg Methods and means for identification of gene features
US9353405B2 (en) 2002-03-12 2016-05-31 Enzo Life Sciences, Inc. Optimized real time nucleic acid detection processes
CA2488858A1 (en) * 2002-06-12 2003-12-24 Research Development Foundation Immunotoxin as a therapeutic agent and uses thereof
CN101538606B (en) * 2009-02-19 2012-03-21 上海浩源生物科技有限公司 Method for detecting one or multiple target nucleic acids and reagent box thereof

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0592626B1 (en) * 1992-03-11 2003-01-29 Dana-Farber Cancer Institute, Inc. METHODS TO CLONE mRNA
US5459037A (en) * 1993-11-12 1995-10-17 The Scripps Research Institute Method for simultaneous identification of differentially expressed mRNAs and measurement of relative concentrations
WO1997029211A1 (en) * 1996-02-09 1997-08-14 The Government Of The United States Of America, Represented By The Secretary, Department Of Health And Human Services RESTRICTION DISPLAY (RD-PCR) OF DIFFERENTIALLY EXPRESSED mRNAs

Also Published As

Publication number Publication date
EP1127159A1 (en) 2001-08-29
CN1331755A (en) 2002-01-16
WO2000026406A1 (en) 2000-05-11
NO20012203L (en) 2001-07-02
KR20010092721A (en) 2001-10-26
MXPA01004550A (en) 2002-09-18
CA2350168A1 (en) 2000-05-11
NO20012203D0 (en) 2001-05-03
JP2002528135A (en) 2002-09-03
AU1108900A (en) 2000-05-22
IL142965A0 (en) 2002-04-21

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