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WO2002031205A3 - Method for rapid subtraction hybridization to isolate a gene which is differentially expressed between two populations of cells - Google Patents

Method for rapid subtraction hybridization to isolate a gene which is differentially expressed between two populations of cells Download PDF

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Publication number
WO2002031205A3
WO2002031205A3 PCT/US2001/031811 US0131811W WO0231205A3 WO 2002031205 A3 WO2002031205 A3 WO 2002031205A3 US 0131811 W US0131811 W US 0131811W WO 0231205 A3 WO0231205 A3 WO 0231205A3
Authority
WO
WIPO (PCT)
Prior art keywords
cdna
pool
double
restriction enzyme
isolate
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/US2001/031811
Other languages
French (fr)
Other versions
WO2002031205A2 (en
Inventor
Paul B Fisher
Hongping Jiang
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.)
Columbia University in the City of New York
Original Assignee
Columbia University in the City of New York
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 Columbia University in the City of New York filed Critical Columbia University in the City of New York
Priority to AU2001296815A priority Critical patent/AU2001296815A1/en
Publication of WO2002031205A2 publication Critical patent/WO2002031205A2/en
Anticipated expiration legal-status Critical
Publication of WO2002031205A3 publication Critical patent/WO2002031205A3/en
Ceased legal-status Critical Current

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

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Analytical Chemistry (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (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)

Abstract

The present invention provides for methods for rapid subtraction hybridization of two pools of double-stranded cDNA to isolate a differentially expressed gene which comprises: isolating RNA from a first and a second population of cells; producing a first and second double-stranded cDNA pool; admixing the first and second pools of cDNA separately with a first restriction enzyme; admixing the digested first pool and second pool of cDNA with an appropriate amount of a double-stranded oligonucleotide; wherein the oligonucleotide sequence comprises (i) an internal recognition site for a second restriction enzyme and (ii) a ligatable cohesive end at one end which is capable of hybridizing with a ligatable cohesive end resulting from digestion of the first pool of cDNA with the first restriction enzyme; performing two separate polymerase chain reactions (PCR) using an aliquot of the ligation mixture (tester cDNA or driver cDNA) admixed with a PCR primer; digesting the PCR products of the tester cDNA with the second restriction enzyme under appropriate digestion conditions; admixing an aliquot of the digestion product (tester cDNA) with an excess of driver cDNA PCR products under hybridization conditions, so as to isolate a differentially expressed gene from among the two pools of double-stranded DNA.
PCT/US2001/031811 2000-10-13 2001-10-11 Method for rapid subtraction hybridization to isolate a gene which is differentially expressed between two populations of cells Ceased WO2002031205A2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU2001296815A AU2001296815A1 (en) 2000-10-13 2001-10-11 Method for rapid subtraction hybridization to isolate a gene which is differentially expressed between two populations of cells

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US68763100A 2000-10-13 2000-10-13
US09/687,631 2000-10-13

Publications (2)

Publication Number Publication Date
WO2002031205A2 WO2002031205A2 (en) 2002-04-18
WO2002031205A3 true WO2002031205A3 (en) 2003-05-08

Family

ID=24761175

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2001/031811 Ceased WO2002031205A2 (en) 2000-10-13 2001-10-11 Method for rapid subtraction hybridization to isolate a gene which is differentially expressed between two populations of cells

Country Status (2)

Country Link
AU (1) AU2001296815A1 (en)
WO (1) WO2002031205A2 (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7251680B2 (en) * 2003-10-31 2007-07-31 Veritas Operating Corporation Single instance backup of email message attachments

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5935788A (en) * 1996-06-25 1999-08-10 Lifespan Biosciences, Inc. Subtractive hybridization techniques for identifying differentially expressed and commonly expressed nucleic acid
US5958738A (en) * 1997-03-24 1999-09-28 Roche Diagnostics Corporation Procedure for subtractive hybridization and difference analysis
US6107023A (en) * 1988-06-17 2000-08-22 Genelabs Technologies, Inc. DNA amplification and subtraction techniques

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6107023A (en) * 1988-06-17 2000-08-22 Genelabs Technologies, Inc. DNA amplification and subtraction techniques
US5935788A (en) * 1996-06-25 1999-08-10 Lifespan Biosciences, Inc. Subtractive hybridization techniques for identifying differentially expressed and commonly expressed nucleic acid
US5958738A (en) * 1997-03-24 1999-09-28 Roche Diagnostics Corporation Procedure for subtractive hybridization and difference analysis

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
BELYAVSKY A ET AL: "PCR-BASED CDNA LIBRARY CONSTRUCTION: GENERAL CDNA LIBRARIES AT THE LEVEL OF A FEW CELLS", NUCLEIC ACIDS RESEARCH, OXFORD UNIVERSITY PRESS, SURREY, GB, vol. 17, no. 8, 1989, pages 2919 - 2932, XP000604679, ISSN: 0305-1048 *
DATABASE EMBL [online] 2 March 2000 (2000-03-02), SCHWER, XP002225709, retrieved from EBI Database accession no. AJ243526 *
JIANG HONGPING ET AL: "RaSH, a rapid subtraction hybridization approach for identifying and cloning differentially expressed genes.", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES, vol. 97, no. 23, 7 November 2000 (2000-11-07), November 7, 2000, pages 12684 - 12689, XP002225708, ISSN: 0027-8424 *
LISITSYN N ET AL: "CLONING THE DIFFERENCES BETWEEN TWO COMPLEX GENOMES", SCIENCE, AMERICAN ASSOCIATION FOR THE ADVANCEMENT OF SCIENCE,, US, vol. 259, no. 5097, 12 February 1993 (1993-02-12), pages 946 - 951, XP000196441, ISSN: 0036-8075 *
SCHWER HEINZ ET AL: "Cloning and characterization of a novel human ubiquitin-specific protease, a homologue of murine UBP43 (Usp18)", GENOMICS, ACADEMIC PRESS, SAN DIEGO, US, vol. 65, no. 1, AJ243526, 1 April 2000 (2000-04-01), pages 44 - 52, XP002153778, ISSN: 0888-7543, Retrieved from the Internet <URL:EBI> *

Also Published As

Publication number Publication date
AU2001296815A1 (en) 2002-04-22
WO2002031205A2 (en) 2002-04-18

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