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US 8,026,050 B1Granted
Methods for modifying an oligonucleotide
AssigneeThird Wave Technologies, Inc.
InventorsJames E. Dahlberg, Mary Ann D. Brow, Victor I. Lyamichev
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Abstract
The present invention relates to means for cleaving a nucleic acid cleavage structure in a site-specific manner. Enzymes, including 5′ nucleases and 3′ exonucleases, are used to detect and identify nucleic acids derived from microorganisms. Methods are provided which allow for the detection and identification of bacterial and viral pathogens in a sample.
Claims6
A method of modifying a polynucleotide, said method comprising:
- (a) providing in combination:
- i) a first oligonucleotide or a molar excess of said first oligonucleotide relative to the concentration of said polynucleotide, with said first oligonucleotide having a 3′ portion capable of reversibly hybridizing to said polynucleotide and a 5′ portion which does not hybridize to said polynucleotide, and
- ii) a 5′-nuclease, and
- (b) reversibly hybridizing under isothermal conditions said polynucleotide and said first oligonucleotide, wherein said first oligonucleotide, when hybridized to said polynucleotide, is cleaved by said 5′-nuclease as a result of the presence of said polynucleotide to provide:
- (i) a first fragment that is substantially non-hybridizable to said polynucleotide, or a first fragment including said 5′ portion and no more than one nucleotide from the 5′ end of said 3′ portion, and (ii) a second fragment that is 3′ of said first fragment with reference to said first oligonucleotide and is substantially hybridizable to said polynucleotide, thereby modifying said polynucleotide.
2 more elements
Dependencies are inferred from the claim text.