WO2014004124A3 - Method for amplification gene fusion complexes - Google Patents

Method for amplification gene fusion complexes Download PDF

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Publication number
WO2014004124A3
WO2014004124A3 PCT/US2013/045864 US2013045864W WO2014004124A3 WO 2014004124 A3 WO2014004124 A3 WO 2014004124A3 US 2013045864 W US2013045864 W US 2013045864W WO 2014004124 A3 WO2014004124 A3 WO 2014004124A3
Authority
WO
WIPO (PCT)
Prior art keywords
nucleic acid
fused
complex
gene fusion
amplification gene
Prior art date
Application number
PCT/US2013/045864
Other languages
French (fr)
Other versions
WO2014004124A2 (en
Inventor
David Scott Johnson
Carolyn G. CONANT
Mathias HOWELL
Original Assignee
Gigagen, 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 Gigagen, Inc. filed Critical Gigagen, Inc.
Publication of WO2014004124A2 publication Critical patent/WO2014004124A2/en
Publication of WO2014004124A3 publication Critical patent/WO2014004124A3/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
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/6853—Nucleic acid amplification reactions using modified primers or templates

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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

This invention is directed to a method for the multiplexed amplification of at least two nucleic acid targets followed by physical linkage of the amplified products. In one aspect of this invention, two distinct nucleic acid target loci are amplified with a common linker sequence that then drives formation of a fused complex. Thermocycling the reaction for more than 50 cycles drives formation of the fused complex. The fused nucleic acid complex can then be assayed by sequencing. This invention has practical application in many biological and medical specialties, such as molecular biology, genetics, oncology, transplantation, and infectious disease.
PCT/US2013/045864 2012-06-14 2013-06-14 Method for amplification gene fusion complexes WO2014004124A2 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201261659581P 2012-06-14 2012-06-14
US61/659,581 2012-06-14

Publications (2)

Publication Number Publication Date
WO2014004124A2 WO2014004124A2 (en) 2014-01-03
WO2014004124A3 true WO2014004124A3 (en) 2014-05-01

Family

ID=49783986

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2013/045864 WO2014004124A2 (en) 2012-06-14 2013-06-14 Method for amplification gene fusion complexes

Country Status (1)

Country Link
WO (1) WO2014004124A2 (en)

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2652155B1 (en) 2010-12-16 2016-11-16 Gigagen, Inc. Methods for massively parallel analysis of nucleic acids in single cells
US9422547B1 (en) 2015-06-09 2016-08-23 Gigagen, Inc. Recombinant fusion proteins and libraries from immune cell repertoires
KR102281923B1 (en) 2017-03-13 2021-07-27 기가젠, 인코포레이티드 Systems and methods for massively parallel combinatorial analysis of single cells
GB201801530D0 (en) 2017-07-07 2018-03-14 Cirrus Logic Int Semiconductor Ltd Methods, apparatus and systems for authentication
EP3941491A4 (en) 2019-03-21 2023-03-29 Gigamune, Inc. Engineered cells expressing anti-viral t cell receptors and methods of use thereof

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
ATANASSOV, IVAN I. ET AL.: "A simple, flexible and efficient PCR-fusion/Gateway cloning procedure for gene fusion, site-directed mutagenesis, short sequence insertion and domain deletions and swaps", PLANT METHODS, vol. 5, no. 14, 2009, pages 1 - 11 *
HORTON, ROBERT M. ET AL.: "Engineering hybrid genes without the use of restriction enzymes: gene splicing by overlap extension", GENE, vol. 77, no. 1, 1989, pages 61 - 68 *
HORTON, ROBERT M. ET AL.: "Gene splicing by overlap extension: tailor-made genes using the polymerase chain reaction", BIOTECHNIQUES, vol. 8, no. 5, 1990, pages 528 - 535 *
WURCH, THIERRY ET AL.: "A modified overlap extension PCR method to create chimeric genes in the absence of restriction enzymes", BIOTECHNOLOGY TECHNIQUES, vol. 12, no. 9, 1998, pages 653 - 657 *
YANG, LIN ET AL.: "Rapid production of gene replacement constructs and generation of a green fluorescent protein-tagged centromeric marker in Aspergillus nidulans", EUKARYOTIC CELL, vol. 3, no. 5, 2004, pages 1359 - 1362 *

Also Published As

Publication number Publication date
WO2014004124A2 (en) 2014-01-03

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