Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1995-04-17
1998-05-12
Jones, W. Gray
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
435 9152, 435 9153, 436 94, 536 2433, 536 2532, 935 77, 935 78, C12Q 168, C07H 2102, C07H 2104, C12P 1934
Patent
active
057503419
ABSTRACT:
Method and compositions are provided for analyzing nucleic acid sequences based on repeated cycles of duplex extension along a single stranded template. Preferably, such extension starts from a duplex formed between an initializing oligonucleotide and the template. The initializing oligonucleotide is extended in an initial extension cycle by ligating an oligonucleotide probe to its end to form an extended duplex. The extended duplex is then repeatedly extended by subsequent cycles of ligation. During each cycle, the identity of one or more nucleotides in the template is determined by a label on, or associated with, a successfully ligated oligonucleotide probe. Preferably, the oligonucleotide probe has a blocking moiety, e.g. a chain-terminating nucleotide, in a terminal position so that only a single extension of the extended duplex takes place in a single cycle. The duplex is further extended in subsequent cycles by removing the blocking moiety and regenerating an extendable terminus. The invention provides a method of sequencing nucleic acids which obviates electrophoretic separation of similarly sized DNA fragments, and which eliminates the difficulties associated with the detection and analysis of spacially overlapping bands of DNA fragments in a gel or like medium. The invention also obviates the need to generate DNA fragments from long single stranded templates with a DNA polymerase.
REFERENCES:
patent: 5002867 (1991-03-01), Macevicz
patent: 5013830 (1991-05-01), Ohtsuka et al.
patent: 5114839 (1992-05-01), Blocker
patent: 5367066 (1994-11-01), Urdea et al.
patent: 5403708 (1995-04-01), Brennan et al.
patent: 5407799 (1995-04-01), Studier
patent: 5503980 (1996-04-01), Cantor
Szybalski, "Proposal for sequencing DNA using ligation of hexamers to generate sequential elongation primers (SPEL-6)," Gene, 90: 177-178 (1990).
Kaczorowski et al, "Assembly of 18-nucleotide primers by ligation of three hexamers: sequencing of large genomes by primer walking," Anal. Biochem., 221: 127-135 (1994).
Hou et al, "Fluorescence-based DNA sequencing with hexamer primers," Nucleic Acids Research, 21: 3331-3332 (1993).
Studier, "A strategy for high-volume sequencing of cosmid DNAs: random and directed priming with a library of oligonucleotides," Proc. Natl. Acad. Sci., 86: 6917-6921 (1989).
Chretien et al, "Multiplex random priming of internal restriction fragments for DNA sequencing," DNA and Cell Biology, 11: 337-343 (1992).
Kotler et al, "DNA sequencing: modular primers assembled from a library of hexamers or pentamers," Proc. Natl. Acad. Sci., 90: 4241-4245 (1993).
Kieleczawa et al, "DNA sequencing by primer walking with strings of contiguous hexamers," Science, 258: 1787-1790 (1992).
Broude et al., Proc. Natl. Acad. Sci. (USA) 91:3072-3076, 1994.
Kuznetsova et al., Molecular Biology (Moscow) 28(2), Part 1: 190-195 (translated from Russia), 1994.
Jones W. Gray
Lynx Therapeutics, Inc.
Macevicz Stephen C.
Powers Vincent M.
Tran Paul B.
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