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1. WO2000039343 - METHOD FOR DETECTING MUTATIONS IN NUCLEIC ACIDS

Publication Number WO/2000/039343
Publication Date 06.07.2000
International Application No. PCT/US1999/031117
International Filing Date 29.12.1999
IPC
C12Q 1/68 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING 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
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
68involving nucleic acids
CPC
C12Q 1/6827
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms
68involving nucleic acids
6813Hybridisation assays
6827for detection of mutation or polymorphism
C12Q 1/6869
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms
68involving nucleic acids
6869Methods for sequencing
Y10T 436/143333
YSECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
10TECHNICAL SUBJECTS COVERED BY FORMER USPC
TTECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
436Chemistry: analytical and immunological testing
14Heterocyclic carbon compound [i.e. , O, S, N, Se, Te, as only ring hetero atom]
142222Hetero-O [e.g., ascorbic acid, etc.]
143333Saccharide [e.g., DNA, etc.]
Applicants
  • CITY OF HOPE [US/US]; 1500 East Duarte Road Duarte, CA 91010-0269, US
Inventors
  • LIU, Qiang; US
  • SOMMER, Steve, S.; US
Agents
  • KERR, Don, M. ; Rothwell, Figg, Ernst & Kurz Suite 701 East 555 13th Street N.W. Columbia Square Washington, DC 20004, US
Priority Data
60/114,57931.12.1998US
Publication Language English (EN)
Filing Language English (EN)
Designated States
Title
(EN) METHOD FOR DETECTING MUTATIONS IN NUCLEIC ACIDS
(FR) PROCEDE DE DETECTION DE MUTATIONS D'ACIDES NUCLEIQUES
Abstract
(EN)
A denaturation fingerprinting method (dnF) involves subjecting a nucleic acid segment of interest to bidirectional cycle sequencing using oppositely oriented primers and incorporating two different dideoxynucleotides (ddNTPs) in the sequencing reaction. The resulting fragments are separated by denaturing electrophoresis. In one embodiment, designated dnF2R, reactions and electrophoretic separation using the two ddNTPs are conducted separately. In an alternative embodiment, designated dnF1R, one of the ddNTPs has a mobility altering modification such that electorphoretic separation occurs when both ddNTPs has are employed in the same reaction. The methods are useful for detecting genetic mutations.
(FR)
L'invention porte sur un procédé de cartographie peptidique dénaturante (dnF) consistant à soumettre un segment d'acide nucléique d'intérêt à un cycle bidirectionnel de séquençage utilisant des amorces de sens opposé tout en incorporant deux didésoxynucléotides (ddNTPs) différents dans la réaction de séquençage. Les fragments résultants sont séparés par électrophorèse dénaturante. Dans l'une des variantes dite dnF2R, les réactions et la séparation par électrophorèse sont effectuées séparément. Dans une autre variante, dite dnF1R, l'un des ddNTPs présente une modification altérant sa mobilité de telle sorte que la séparation par électrophorèse intervient lorsque les deux ddNTPs participent à la même réaction. Ces procédés permettent de détecter des mutations génétiques.
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