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WO2000008208A3 - Reverse transcription and amplification processes and primers therefor - Google Patents

Reverse transcription and amplification processes and primers therefor Download PDF

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Publication number
WO2000008208A3
WO2000008208A3 PCT/GB1999/002579 GB9902579W WO0008208A3 WO 2000008208 A3 WO2000008208 A3 WO 2000008208A3 GB 9902579 W GB9902579 W GB 9902579W WO 0008208 A3 WO0008208 A3 WO 0008208A3
Authority
WO
WIPO (PCT)
Prior art keywords
heeled
reverse transcription
primers
amplification processes
primers therefor
Prior art date
Legal status (The legal status 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 status listed.)
Ceased
Application number
PCT/GB1999/002579
Other languages
French (fr)
Other versions
WO2000008208A2 (en
Inventor
Thomas Charles Freeman
Peter John Richardson
Alistair K Dixon
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Medical Research Council
Original Assignee
Medical Research Council
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 Medical Research Council filed Critical Medical Research Council
Priority to AU52943/99A priority Critical patent/AU5294399A/en
Publication of WO2000008208A2 publication Critical patent/WO2000008208A2/en
Publication of WO2000008208A3 publication Critical patent/WO2000008208A3/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • 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
    • C12Q1/6853—Nucleic acid amplification reactions using modified primers or templates
    • 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
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/158—Expression markers

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Immunology (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Analytical Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Biotechnology (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)

Abstract

A method is provided for the expression profiling of single cells. The method employs a first heeled primer for reverse transcription of mRNA in a sample to provide first strand cDNA species, and then a second heeled primer population to generate second strand cDNAs. The non-heeled portion of the second heeled primers are capable of hybridizing to the reverse transcribed first strands of cDNA species, at least one along the lengths thereof. Due to the presence of random and preselected sequences in the second primers a qualitatively more uniform and therefore representative cDNA profile is produced from cellular mRNAs.
PCT/GB1999/002579 1998-08-05 1999-08-05 Reverse transcription and amplification processes and primers therefor Ceased WO2000008208A2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU52943/99A AU5294399A (en) 1998-08-05 1999-08-05 Reverse transcription and amplification processes and primers therefor

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB9817055.8 1998-08-05
GBGB9817055.8A GB9817055D0 (en) 1998-08-05 1998-08-05 Reverse transcription and amplification processes and primers therefore

Publications (2)

Publication Number Publication Date
WO2000008208A2 WO2000008208A2 (en) 2000-02-17
WO2000008208A3 true WO2000008208A3 (en) 2000-05-11

Family

ID=10836764

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/GB1999/002579 Ceased WO2000008208A2 (en) 1998-08-05 1999-08-05 Reverse transcription and amplification processes and primers therefor

Country Status (3)

Country Link
AU (1) AU5294399A (en)
GB (1) GB9817055D0 (en)
WO (1) WO2000008208A2 (en)

Cited By (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6946251B2 (en) 2001-03-09 2005-09-20 Nugen Technologies, Inc. Methods and compositions for amplification of RNA sequences using RNA-DNA composite primers
US7094536B2 (en) 2001-03-09 2006-08-22 Nugen Technologies, Inc. Methods and compositions for amplification of RNA sequences
US7294461B2 (en) 2000-06-26 2007-11-13 Nugen Technologies, Inc. Methods and compositions for transcription-based nucleic acid amplification
US7402386B2 (en) 2003-04-14 2008-07-22 Nugen Technologies, Inc. Global amplification using random priming by a composite primer
US7771934B2 (en) 2000-12-13 2010-08-10 Nugen Technologies, Inc. Methods and compositions for generation of multiple copies of nucleic acid sequences and methods of detection thereof
US7846733B2 (en) 2000-06-26 2010-12-07 Nugen Technologies, Inc. Methods and compositions for transcription-based nucleic acid amplification
US7846666B2 (en) 2008-03-21 2010-12-07 Nugen Technologies, Inc. Methods of RNA amplification in the presence of DNA
US7939258B2 (en) 2005-09-07 2011-05-10 Nugen Technologies, Inc. Nucleic acid amplification procedure using RNA and DNA composite primers
US8034568B2 (en) 2008-02-12 2011-10-11 Nugen Technologies, Inc. Isothermal nucleic acid amplification methods and compositions
US8143001B2 (en) 2003-12-29 2012-03-27 Nugen Technologies, Inc. Methods for analysis of nucleic acid methylation status and methods for fragmentation, labeling and immobilization of nucleic acids
US9206418B2 (en) 2011-10-19 2015-12-08 Nugen Technologies, Inc. Compositions and methods for directional nucleic acid amplification and sequencing
US9650628B2 (en) 2012-01-26 2017-05-16 Nugen Technologies, Inc. Compositions and methods for targeted nucleic acid sequence enrichment and high efficiency library regeneration
US9745614B2 (en) 2014-02-28 2017-08-29 Nugen Technologies, Inc. Reduced representation bisulfite sequencing with diversity adaptors
US9822408B2 (en) 2013-03-15 2017-11-21 Nugen Technologies, Inc. Sequential sequencing
US9957549B2 (en) 2012-06-18 2018-05-01 Nugen Technologies, Inc. Compositions and methods for negative selection of non-desired nucleic acid sequences
US11028430B2 (en) 2012-07-09 2021-06-08 Nugen Technologies, Inc. Methods for creating directional bisulfite-converted nucleic acid libraries for next generation sequencing

Families Citing this family (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU6695600A (en) * 1999-07-19 2001-02-05 Cambridge University Technical Services Limited A method for amplifying low abundance nucleic acid sequences and means for performing said method
US6692918B2 (en) * 1999-09-13 2004-02-17 Nugen Technologies, Inc. Methods and compositions for linear isothermal amplification of polynucleotide sequences
CA2431170C (en) * 2000-12-12 2011-07-26 Aisin Seiki Kabushiki Kaisha Method of analyzing expression of gene
EP1275738A1 (en) * 2001-07-11 2003-01-15 Roche Diagnostics GmbH Method for random cDNA synthesis and amplification
US7414111B2 (en) 2001-09-19 2008-08-19 Alexion Pharmaceuticals, Inc. Engineered templates and their use in single primer amplification
CA2460639C (en) 2001-09-19 2013-07-16 Alexion Pharmaceuticals, Inc. Engineered templates and their use in single primer amplification
CA2478875A1 (en) 2002-03-11 2003-09-25 Nugen Technologies, Inc. Methods for generating double stranded dna comprising a 3' single stranded portion and uses of these complexes for recombination
GB0216026D0 (en) * 2002-07-10 2002-08-21 Cambridge Biotechnology Ltd Nucleic acid amplification method
EP3068883B1 (en) 2013-11-13 2020-04-29 Nugen Technologies, Inc. Compositions and methods for identification of a duplicate sequencing read
CA2957633A1 (en) 2014-08-06 2016-02-11 Nugen Technologies, Inc. Digital measurements from targeted sequencing
US12492430B2 (en) 2017-04-11 2025-12-09 Tecan Genomics, Inc. Library quantitation and qualification
US11099202B2 (en) 2017-10-20 2021-08-24 Tecan Genomics, Inc. Reagent delivery system
US12059674B2 (en) 2020-02-03 2024-08-13 Tecan Genomics, Inc. Reagent storage system

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991015601A1 (en) * 1990-04-05 1991-10-17 The United States Of America, As Represented By The Secretary, U.S. Department Of Commerce Modified rna template-specific polymerase chain reaction
WO1998008973A1 (en) * 1996-08-29 1998-03-05 Cancer Research Campaign Technology Limited Global amplification of nucleic acids

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991015601A1 (en) * 1990-04-05 1991-10-17 The United States Of America, As Represented By The Secretary, U.S. Department Of Commerce Modified rna template-specific polymerase chain reaction
WO1998008973A1 (en) * 1996-08-29 1998-03-05 Cancer Research Campaign Technology Limited Global amplification of nucleic acids

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
CHEUNG AND NELSON: "WHOLE GENOME AMPLIFICATION USING A DEGENERATE OLIGONUCLEOTIDE PRIMER ALLOWS HUNDREDS OF GENOTYPES TO BE PERFORMED ON LESS THAN ONE NANOGRAM OF GENOMIC DNA", PNAS, vol. 93, 1996, pages 14676 - 14679, XP002128316 *
DIXON ET AL.: "EXPRESSION PROFILING OF SINGLE CELLS USING 3 PRIME END AMPLIFICATION (TPEA) PCR", NUCLEIC ACIDS RESEARCH, vol. 26, no. 19, October 1998 (1998-10-01), pages 4426 - 4431, XP002128317 *
FROUSSARD: "A RANDOM-PCR METHOD (rPCR) TO CONSTRUCT WHOLE cDNA LIBRARY FROM LOW AMOUNTS OF RNA", NUCLEIC ACIDS RESEARCH, vol. 20, no. 11, 1992, pages 2900, XP002128315 *
TELENIUS H ET AL: "DEGENERATE OLIGONUCLEOTIDE-PRIMED PCR: GENERAL AMPLIFICATION OF TARGET DNA BY A SINGLE DEGENERATE PRIMER", GENOMICS,US,ACADEMIC PRESS, SAN DIEGO, vol. 13, 1992, pages 718-725, XP000199116, ISSN: 0888-7543 *
ZHAO ET AL.: "3'-END cDNA POOL SUITABLE FOR DIFFERENTIAL DISPLAY FROM A SMALL NUMBER OF CELLS", BIOTECHNIQUES, vol. 24, May 1998 (1998-05-01), pages 842 - 852, XP002128314 *
ZHAO ET AL.: "NEW PRIMER STRATEGY IMPROVES PRECISION OF DIFFERENTIAL DISPLAY", BIOTECHNIQUES, vol. 18, no. 5, 1995, pages 842 - 850, XP002128267 *

Cited By (27)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7846733B2 (en) 2000-06-26 2010-12-07 Nugen Technologies, Inc. Methods and compositions for transcription-based nucleic acid amplification
US7294461B2 (en) 2000-06-26 2007-11-13 Nugen Technologies, Inc. Methods and compositions for transcription-based nucleic acid amplification
US7771934B2 (en) 2000-12-13 2010-08-10 Nugen Technologies, Inc. Methods and compositions for generation of multiple copies of nucleic acid sequences and methods of detection thereof
US8334116B2 (en) 2000-12-13 2012-12-18 Nugen Technologies, Inc. Methods and compositions for generation of multiple copies of nucleic acid sequences and methods of detection thereof
US7351557B2 (en) 2001-03-09 2008-04-01 Nugen Technologies, Inc. Amplification of RNA sequences using composite RNA-DNA primers and strand displacement
US7771946B2 (en) 2001-03-09 2010-08-10 Nugen Technologies, Inc. Methods, kits and compositions for single primer linear isothermal amplification of nucleic acid sequences
US7094536B2 (en) 2001-03-09 2006-08-22 Nugen Technologies, Inc. Methods and compositions for amplification of RNA sequences
US9181582B2 (en) 2001-03-09 2015-11-10 Nugen Technologies, Inc. Compositions for amplification of RNA sequences using composite primers
US8071311B2 (en) 2001-03-09 2011-12-06 Nugen Technologies, Inc. Methods and compositions for amplification of RNA sequences
US6946251B2 (en) 2001-03-09 2005-09-20 Nugen Technologies, Inc. Methods and compositions for amplification of RNA sequences using RNA-DNA composite primers
US7402386B2 (en) 2003-04-14 2008-07-22 Nugen Technologies, Inc. Global amplification using random priming by a composite primer
US9175325B2 (en) 2003-04-14 2015-11-03 Nugen Technologies, Inc. Global amplification using a randomly primed composite primer
US8465950B2 (en) 2003-04-14 2013-06-18 Nugen Technologies, Inc. Global amplification using a randomly primed composite primer
US8143001B2 (en) 2003-12-29 2012-03-27 Nugen Technologies, Inc. Methods for analysis of nucleic acid methylation status and methods for fragmentation, labeling and immobilization of nucleic acids
US8852867B2 (en) 2005-09-07 2014-10-07 Nugen Technologies, Inc. Nucleic acid amplification procedure using RNA and DNA composite primers
US7939258B2 (en) 2005-09-07 2011-05-10 Nugen Technologies, Inc. Nucleic acid amplification procedure using RNA and DNA composite primers
US8034568B2 (en) 2008-02-12 2011-10-11 Nugen Technologies, Inc. Isothermal nucleic acid amplification methods and compositions
US7846666B2 (en) 2008-03-21 2010-12-07 Nugen Technologies, Inc. Methods of RNA amplification in the presence of DNA
US9206418B2 (en) 2011-10-19 2015-12-08 Nugen Technologies, Inc. Compositions and methods for directional nucleic acid amplification and sequencing
US10036012B2 (en) 2012-01-26 2018-07-31 Nugen Technologies, Inc. Compositions and methods for targeted nucleic acid sequence enrichment and high efficiency library generation
US9650628B2 (en) 2012-01-26 2017-05-16 Nugen Technologies, Inc. Compositions and methods for targeted nucleic acid sequence enrichment and high efficiency library regeneration
US10876108B2 (en) 2012-01-26 2020-12-29 Nugen Technologies, Inc. Compositions and methods for targeted nucleic acid sequence enrichment and high efficiency library generation
US9957549B2 (en) 2012-06-18 2018-05-01 Nugen Technologies, Inc. Compositions and methods for negative selection of non-desired nucleic acid sequences
US11028430B2 (en) 2012-07-09 2021-06-08 Nugen Technologies, Inc. Methods for creating directional bisulfite-converted nucleic acid libraries for next generation sequencing
US9822408B2 (en) 2013-03-15 2017-11-21 Nugen Technologies, Inc. Sequential sequencing
US10760123B2 (en) 2013-03-15 2020-09-01 Nugen Technologies, Inc. Sequential sequencing
US9745614B2 (en) 2014-02-28 2017-08-29 Nugen Technologies, Inc. Reduced representation bisulfite sequencing with diversity adaptors

Also Published As

Publication number Publication date
GB9817055D0 (en) 1998-09-30
WO2000008208A2 (en) 2000-02-17
AU5294399A (en) 2000-02-28

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