WO2009117698A3 - Methods of rna amplification in the presence of dna - Google Patents

Methods of rna amplification in the presence of dna Download PDF

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Publication number
WO2009117698A3
WO2009117698A3 PCT/US2009/037870 US2009037870W WO2009117698A3 WO 2009117698 A3 WO2009117698 A3 WO 2009117698A3 US 2009037870 W US2009037870 W US 2009037870W WO 2009117698 A3 WO2009117698 A3 WO 2009117698A3
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WO
WIPO (PCT)
Prior art keywords
methods
dna
rna
amplification
marked
Prior art date
Application number
PCT/US2009/037870
Other languages
French (fr)
Other versions
WO2009117698A2 (en
Inventor
Nurith Kurn
Original Assignee
Nugen Technologies, Inc.
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 Nugen Technologies, Inc. filed Critical Nugen Technologies, Inc.
Priority to GB1014134.9A priority Critical patent/GB2470672B/en
Publication of WO2009117698A2 publication Critical patent/WO2009117698A2/en
Publication of WO2009117698A3 publication Critical patent/WO2009117698A3/en

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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
    • C12N15/1096—Processes for the isolation, preparation or purification of DNA or RNA cDNA Synthesis; Subtracted cDNA library construction, e.g. RT, RT-PCR
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • 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
    • C12Q1/6809—Methods for determination or identification of nucleic acids involving differential detection
    • 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
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/6848—Nucleic acid amplification reactions characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction

Abstract

The invention provides methods for amplification of RNA The methods are particularly suitable for specifically amplifying RNA in the presence of DNA The methods involve producing a marked first primer extension product from a target RNA in the presence of a DNA-dependent DNA polymerase inhibitor, which prevents replication of DNA by the reverse transcriptase enzyme The marked nucleic acid products are subsequently selectively amplified in the presence on non-marked nucleic acids The methods are useful for production and analysis of polynucleotide sequences complementary to an RNA sequence The methods are useful for preparation of nucleic acid libraries and substrates for analysis of gene expression of cells in biological samples The invention also provides compositions and kits for practicing the amplification methods, as well as methods which use the amplification products.
PCT/US2009/037870 2008-03-21 2009-03-20 Methods of rna amplification in the presence of dna WO2009117698A2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
GB1014134.9A GB2470672B (en) 2008-03-21 2009-03-20 Methods of RNA amplification in the presence of DNA

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
US3869608P 2008-03-21 2008-03-21
US61/038,696 2008-03-21
US7499108P 2008-06-23 2008-06-23
US61/074,991 2008-06-23
US8581108P 2008-08-01 2008-08-01
US61/085,811 2008-08-01

Publications (2)

Publication Number Publication Date
WO2009117698A2 WO2009117698A2 (en) 2009-09-24
WO2009117698A3 true WO2009117698A3 (en) 2009-12-10

Family

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Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2009/037870 WO2009117698A2 (en) 2008-03-21 2009-03-20 Methods of rna amplification in the presence of dna

Country Status (3)

Country Link
US (1) US7846666B2 (en)
GB (1) GB2470672B (en)
WO (1) WO2009117698A2 (en)

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