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AssigneeNew England Biolabs, Inc.
InventorsWilliam E. Jack, Ira Schildkraut, Julie Forney Menin
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The present invention relates to the use of site-specific nucleic acid nicking enzymes to create single-stranded regions in duplex nucleic acids. Such single-stranded regions can take the form of gaps interior to the duplex, or terminal single-stranded regions. Single-stranded termini can be crafted to allow linkage of various elements via base-pairing with elements containing a complementary single-stranded region. This joining is useful, for example, in an ordered, oriented assembly of DNA modules to create cloning or expression vectors. This joining is also useful in attaching detection probes and purifying DNA molecules containing the single-stranded region. Gaps are useful in similar applications, including attaching detection or purification probes.
A method for creating target single strand regions in a plurality of double stranded DNA molecules for use in joining the DNA molecules, comprising:
A method for creating a target single strand region at a terminus of a linear double stranded DNA molecule for use in joining the DNA molecule to a second DNA molecule by means of the single strand region, or for detecting, purifying or selectively mutagenizing the DNA molecule, comprising
A method for creating target single strand regions in a double stranded DNA molecule for use in detecting, purifying or selectively mutagenizing the DNA molecule, the method comprising:
Dependencies are inferred from the claim text.